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 Contents

Preface................................................................. ix 15. Non-spore-forming anaerobes...............205

Acknowledgements............................................ xi 16. Clostridium species.................................. 215
17. Enterobacteriaceae.....................................239
18. Pseudomonas, Burkholderia and
SECTION 1: General procedures Stenotrophomonas species........................275
in microbiology 19. Aeromonas, Plesiomonas and
Vibrio species...........................................289
1. Collection and submission of
20. Actinobacillus species...............................297
diagnostic specimens.................................. 3
21. Pasteurella, Mannheimia, Bibersteinia
2. Bacterial pathogens: Microscopy,
and Avibacterium species.........................307
culture and identification........................... 9
22. Francisella tularensis................................. 317
3. Serological diagnosis................................49
23. Brucella species........................................325
4. Molecular techniques in diagnostic
24. Campylobacter, Arcobacter and
microbiology . ..........................................59
Helicobacter species.................................335
5. The isolation of viruses and
25. Lawsonia intracellularis............................345
the detection of virus and .
26. Haemophilus and Histophilus
viral antigens.............................................67
species......................................................349
6. Antimicrobial agents................................79
27. Taylorella species......................................355
28. Bordetella species.....................................359
SECTION 2: Bacteriology 29. Moraxella species.....................................369
30. Glucose non-fermenting,
7. Staphylococcus species.............................. 105 Gram-negative bacteria...........................375
8. The streptococci and related cocci......... 121 31. The spirochaetes..................................... 381
9. Corynebacterium species and 32. Miscellaneous Gram-negative
Rhodococcus equi .....................................135 bacteria....................................................399
10. The Actinobacteria..................................147 33. Chlamydiales.............................................407
11. Mycobacterium species............................. 161 34. Rickettsiales and Coxiella burnetii............ 417
12. Listeria species.........................................177 35. The mycoplasmas
13. Erysipelothrix species................................187 (class: Mollicutes)...................................423
14. Bacillus species........................................195 36. Mastitis ...................................................433

vii
 Contents

56. Birnaviridae.............................................. 613

SECTION 3: Mycology 57. Flaviviridae............................................... 617
37. Introduction to the pathogenic 58. Arteriviridae..............................................629
fungi.........................................................457 59. Togaviridae................................................635
38. The dermatophytes................................. 471 60. Orthomyxoviridae......................................639
39. Aspergillus species and 61. Paramyxoviridae........................................645
Pneumocystis carinii.................................. 481 62. Coronaviridae............................................655
40. The pathogenic yeasts.............................487 63. Rhabdoviridae...........................................665
41. The dimorphic fungi..............................497 64. Bunyaviridae.............................................673
42. The pathogenic Zygomycetes...................505 65. Retroviridae...............................................679
43. Fungi causing subcutaneous 66. Bornaviridae............................................. 691
mycoses................................................... 513 67. Prions (proteinaceous infectious
44. Mycotoxins and mycotoxicoses............. 521 agents).....................................................693

SECTION 5: Zoonoses
SECTION 4: Virology
(including prions) 68. Zoonoses.................................................703

45. Parvoviridae.............................................. 541

46. Circoviridae...............................................547 SECTION 6: A systems approach
47. Papillomaviridae....................................... 551 to infectious diseases on
48. Adenoviridae.............................................555 a species basis
49. Herpesviridae............................................559
50. Asfarviridae...............................................575 69. Infectious diseases..................................735
51. Poxviridae.................................................579
52. Picornaviridae...........................................587 Appendix 1 Reagents and stains...................845
53. Caliciviridae..............................................597 Appendix 2 Culture and transport media.... 851
54. Astroviridae...............................................603
55. Reoviridae.................................................605 Index................................................................857

viii
 Preface

A lot of water has passed under the bridge since the popular first edition of this book. Needless to
say the writing of textbooks is not a full-time occupation for any of the authors but rather a com-
plementary activity to teaching, research and diagnostic service. This project was driven by a love
of books. On more than one occasion the writing faltered due to other more pressing commitments
and the vicissitudes of life in general. We have tried to retain the essential structure and character
of the first edition while updating and adding new material as necessary. In particular, the virology
section has been greatly expanded with the addition of a chapter for each of the families containing
viruses of veterinary importance. For taxonomy we have relied heavily on resources such as the List
of Prokaryotic Names with Standing in Nomenclature (www.bacterio.cict.fr/), the virus list of the
International Committee on Taxonomy of Viruses (ictvonline.org/index.asp) and Index Fungorum
(www.indexfungorum.org/). For information on the diagnosis of many of the more important
infectious diseases of farm animals we have frequently consulted the Manual of Diagnostic Tests
and Vaccines for Terrestrial Animals on the website (www.oie.int/en/international-standard-setting/
terrestrial-manual/access-online/) of the OIE (Office International des Epizooties – World Organisa-
tion for Animal Health). We hope we have succeeded in producing a book that is useful, informative
and appealing.

Dublin, 2013

ix
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 Acknowledgements

Assembling material for a book requires forward planning, attention to detail and the means to put
in place the knowledge and ideas of the authors. We were fortunate to have the resources of the
first edition to draw upon in bringing this project to fruition. We are indebted to Professor Emeritus
Joe Quinn for being our mentor and greatest supporter in this endeavour.
Veterinary microbiology lends itself to illustration and we have tried to include suitable illustrative
material in the form of colour images and line drawings. A number of colleagues supplied images
which we would like to acknowledge: Professor KP Baker (38.7), Centres for Disease Control (31.13
and 34.2), Dr SJ Cook and Professor CJ Issel (3.3, 3.16 and 3.22), Dr DO Cordes (11.14, 42.7, 42.8,
42.9 and 44.7), Professor FWG Hill (29.1), Dr L Hoffmann (31.12), Dr G Joseph (18.7, 18.8 and
22.1), Professor JF Kazda (11.5, 11.6 and 11.7), Mr Aonghus Lane (36.21 and 36.22) and Dr N
Seiranganathan (35.8).
We also wish to acknowledge our predecessors in the Dublin Vet School, particularly Mr BT
Whitty, Mr MA Gallaher, Professor PJ Quinn and Dr Margery Carter who were prominent in our
formative training and who left for posterity a wealth of illustrative material and stained smears,
many of which we have used to illustrate individual chapters. We would also like to record our
appreciation to colleagues who furnished material for photography: Dr HF Bassett (10.17), Mr MJ
Casey (68.4), Mr RP Cooney (11.12), Mr P Costigan (5.4, 5.5, 51.1 and 51.2), Dr WJC Donnelly
(12.2, 12.3 and 67.2), Mr E Fitzpatrick (41.2), Dr HA Larkin (35.2), Dr GHK Lawson (33.4), Mr JB
Power (31.8 and 31.10), Dr GR Scott (34.1 and 35.1) and Professor Emeritus BJ Sheahan (39.2).
Finally, the authors would like to record their appreciation to the publishers for their patience
and encouragement while the book was in preparation, in particular Joyce Rodenhuis, Rita Deme-
triou-Swanwick, Louisa Welch, Robert Edwards, Veronika Watkins and Clive Hewat.

xi
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 Section 1
General procedures in microbiology

1
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 Chapter
Collection and submission of
diagnostic specimens
Choosing and Working with
a Laboratory
In choosing a laboratory clinicians must consider the cost
of the service and the ease of access. However, it is equally
important to determine if a laboratory is accredited, has a
specialization in the relevant species, offers a service at
weekends and holidays and has staff with the necessary
expertise and training to assist with disease investigations.
The quality of the dialogue between the veterinary practice
and the laboratory has an enormous influence on the
effectiveness of the service. Good communication is essen-
tial. The clinician and the microbiologist must establish
and maintain contact with each other. An accurate diag-
nosis is based on the interpretation of both clinical and
laboratory data.
If the clinician is not absolutely certain of the correct
sample to collect they should seek advice from the labora-
tory prior to collecting the samples. The samples must be
appropriate for the purpose required which may be diag-
nosis, certification, surveillance or the monitoring of
vaccine efficacy or the response to antimicrobial therapy.
A clinical diagnosis cannot be confirmed by the examina-
tion of inappropriate samples. In disease control situa-
tions samples must be collected systematically for a
definite purpose, for example, the lifting of movement
restrictions or the commencement of breeding. Laboratory
tests can be both expensive and time-consuming. It is
important for all concerned that resources are not wasted
in the generation of irrelevant information.
Both the veterinary practitioner and the laboratory have
an essential role in the isolation of new agents and the
detection of emerging disease patterns. In influenza out-
breaks, for example, the submission of samples to the
laboratory for virus isolation is necessary to identify new
© 2013 Elsevier Ltd
1
strains of viruses. This facilitates the updating of vaccines
with epidemiologically relevant viruses.
Some laboratories are involved in contract research and
perform safety and efficacy studies for the regulatory
authorities. All diagnostic laboratories have a role to play
in the monitoring of the efficacy of existing products. Cli-
nicians have a responsibility to inform the microbiologist
of the vaccination and/or therapeutic history of the animal
to facilitate independent assessment of different products
and treatment regimens. Microbiologists in turn have a
responsibility to advise clinicians with regard to their
findings.
General Principles for
Sample Collection
• Samples should be taken from the affected site(s) as
early as possible following the onset of clinical signs.
This is particularly important in viral diseases as
shedding of virus is usually maximal early in the
infection. This is also true of enteric bacterial
pathogens.
• It is useful to collect samples from clinical cases and
in-contact animals, particularly if there has been
an outbreak of disease. In-contact animals may
be at an earlier stage in the infection with a greater
chance of them shedding substantial numbers of
microorganisms.
• Samples should be obtained from the edge of lesions
and some macroscopically normal tissue included.
Microbial replication will be most active at the
lesion’s edge.
• It is important to collect specimens as aseptically as
possible, otherwise the relevant pathogen may be
overgrown by numerous contaminating bacteria
3
Section | 1 | General Procedures in Microbiology
Figure 1.1 Grossly contaminated blood agar plate
demonstrating the need to collect specimens as carefully as
possible.
(Fig. 1.1). In certain circumstances a guarded swab
should be used to bypass an area with a large
population of normal flora.
• The laboratory should be informed if treatment has
commenced in order that counteractive measures
may be taken to increase the possibility of isolating
bacteria or that an alternative method of detection
such as polymerase chain reaction (PCR) may be
employed.
• When possible a generous amount of sample should
be taken and submitted, such as blocks of tissue
(approximately 2 cm3), biopsy material, or several
millilitres of pus, exudate or faeces. For serology, at
least 5 mL of blood should be obtained to allow a
number of tests to be carried out if necessary and to
allow the sample to be stored and tested with
subsequent samples.
• Cross-contamination between samples must be
avoided. This is essential where a highly sensitive
amplification technique such as PCR is to be used
for the detection of the aetiological agent.
• Precautions must be taken to avoid human infection
where a zoonotic condition is suspected.
Tissue
Postmortem material should be collected as soon as pos-
sible after death. However it is sometimes worthwhile
submitting old and even partially decomposed samples if
that is all that is available. In the outbreak of African horse
sickness in the Iberian Peninsula in the late 1980s virus
was isolated from the bone marrow of a horse that had
been buried for eight days. An organism does not have to
be viable or even entire for its genome to be detected by
polymerase chain reaction. Thus it may be possible to
obtain a diagnosis by PCR from a sample that is totally
unsuitable for histopathological examination or agent
isolation.
4
Figure 1.2 Sterile disposable containers and swabs for
specimen collection.
Tissues from outside the body cavities should be col-
lected first followed by tissues from the thorax and then
the abdomen. Sterile instruments should be used to collect
tissue samples of at least 1 cm3 which should then be
placed in separate sterile screw-capped jars (Fig. 1.2). If
the laboratory is some distance away tissue may be for-
warded in virus or bacterial transport medium. It is impor-
tant to remember that virus transport medium usually
contains antibiotics thus rendering the sample unsuitable
for bacterial examination. Tissue for histopathological
examination should be placed in at least 10 times its
volume of neutral buffered 10% formalin.
In cases of abortion the whole foetus and placenta
should be submitted. If this is not possible then samples
of tissue, a piece of affected placenta, foetal abomasal
contents (ruminants) and uterine discharge (if applicable)
should be forwarded to the laboratory. A clotted blood
sample from the dam for serological examination may
yield additional information.
Swabs and discharges
Fluids are preferable to swabs as the greater sample volume
increases the likelihood of detecting the causal organism.
Samples for agent isolation should be placed in sterile
containers. Viruses and many bacteria are susceptible to
desiccation especially if collected on a dry swab. Formulae
for suitable transport media for viruses, chlamydia and
other organisms are given in Appendix 2. Whenever pos-
sible the sample should be collected from the specific site
of infection. The usual short cotton wool swabs are gener-
ally unsatisfactory for obtaining nasopharyngeal speci-
mens of epithelial cells and mucus for the investigation of
respiratory disease of large animals. Guarded swabs are
necessary for certain bacteriological examinations where
misleading results could be generated due to contami-
nants from adjacent sites that are colonized with bacterial
flora. Similarly, fungal organisms from the environment
 Collection and submission of diagnostic specimens
readily contaminate the nasal passages and upper trachea.
The diagnostic laboratory should be consulted before
collecting samples for the isolation of specific pathogens
that require specialized media or culture conditions, for
example, Taylorella equigenitalis, Chlamydophila psittaci or
Mycoplasma species. The laboratory will either supply spe-
cialist swabs and transport media or recommend a repu-
table source, as appropriate.
Samples from skin lesions
If intact pustules or vesicles are present, the surface
should be disinfected with 70% ethyl alcohol, allowed to
dry, and material aspirated from the lesion with a sterile
syringe and fine needle. A swab may be taken from the
raw surface of ulcers. A biopsy of wound tissue should be
collected after the superficial area has been cleaned and
debrided.
In cases where ringworm is suspected, hair should be
plucked from the lesion and the edge of the lesion scraped
with a blunt scalpel blade until blood begins to ooze.
Plucked hair, skin scrapings (including the scalpel blade
itself) and any scab material that is present should be
submitted. These specimens will also allow detection of
mange or a bacterial infection, if present. In cases of orf
the crust and scrapings from the edge of the lesion should
be collected. In birds feather follicle skin is useful in the
diagnosis of Marek’s disease.
Blood
Blood should be withdrawn using an aseptic technique
into a dry syringe or vacutainer. If collected in a syringe,
care must be taken not to cause haemolysis. The needle
should be removed prior to expelling the sample carefully
into a sterile dry tube. It is not acceptable to submit blood
to the laboratory in a syringe. Glass tubes are fragile.
However, blood clots often retract poorly in plastic bottles
making it difficult to separate the serum. Whole blood
should never be frozen prior to submission to a laboratory
as the ensuing lysis of red blood cells makes it impossible
to perform many serological assays.
Serological tests are usually performed on the serum
harvested from clotted samples. Paired samples are fre-
quently required to make a definitive diagnosis on the
basis of serology. It is advisable to confirm the most appro-
priate sampling interval with the microbiologist who
should also advise as to usefulness of a single blood
sample.
Blood for the isolation of viruses or bacteria should be
prevented from clotting. The laboratory should advise on
the most appropriate anticoagulant. As a bacteraemia can
be intermittent, it is advisable to take more than one
sample within a 24-hour period. The blood should be
added aseptically and without delay to one of the special
commercial blood-culture bottles. Blood samples for
Chapter |1|
culture should be kept cool and submitted to the labora-
tory as quickly as possible.
Faeces
A faeces sample freshly voided or collected from the
rectum is preferable to a rectal swab which often does not
have enough faecal matter for agent detection. A faeces
sample (about the size of the end of a thumb) may be
forwarded to the laboratory without transport medium.
Faecal swabs should be placed in medium such as buffered
glycerol saline to avoid desiccation. Some organisms are
shed intermittently and samples may need to be collected
over several days.
Urine samples
Urine samples may be submitted for urinalysis, bacterial
microscopy and culture or for a bacterial viable count to
establish whether clinical bacteriuria is present. For bac­
teriological procedures the preferred methods of collec-
tion are by cystocentesis, by catheter or mid-stream urine
sample.
Abscesses
If possible about 3 mL of pus should be collected together
with scrapings from the wall of the abscess. Pus at the
centre of an abscess is often sterile. Pus from recently
formed abscesses will yield the best cultural results. Anaer-
obic bacteria can often be cultured from abscesses.
Eye
A conjunctival swab may be taken gently holding the
palpebrae apart. Scrapings may also be taken with a fine
sterile spatula. The cells should be washed carefully into
transport medium.
Bovine mastitic milk samples
Milk samples should be collected from cows as soon as
possible after the mastitis is first noticed and not from
animals treated with either intramammary or systemic
antibiotics. The udder should not be rinsed with water
unless very dirty. If the udder and teats are washed, they
should be dried thoroughly with a paper towel. Usually
it is sufficient to wipe the teats vigorously, using 70%
ethyl alcohol on cotton wool, paying special attention
to the teat sphincters. Antiseptics should be avoided. The
two teats furthest from the operator are wiped first and
then the two nearest teats. The sterile narrow-necked col-
lection bottle must be held almost horizontally. The first
squirt of milk from each teat is discarded and then, for a
composite sample, a little milk from each quarter is
5
Section | 1 | General Procedures in Microbiology
directed into the bottle. The milk should be collected from
the two near teats first and then from the two far teats, so
that one’s arm is less likely to accidentally brush against a
cleaned teat.
Specimens for anaerobic culture
A good collection method is essential because many anaer-
obes do not survive frank exposure to the oxygen in the
air for more than 20 minutes. It is important not to con-
taminate the samples by contact with adjacent mucosal
surfaces as these have a resident anaerobic flora. Speci-
mens from animals that have been dead for more than
four hours are usually unsuitable because of the rapid
postmortem invasion of the animal body by anaerobes
from the intestinal tract. Bone marrow is a good specimen
to collect for the diagnosis of blackleg or malignant
oedema as bone marrow appears to be one of the last
tissues to be invaded by contaminating bacteria. A piece
of rib stripped of the periosteum could be submitted to
the laboratory for the extraction of bone marrow. Any
specimens for the attempted isolation of anaerobes must
arrive at the laboratory as soon as possible after collection.
Collection of samples for anaerobe culture on ordinary
swabs is usually of no value. Acceptable samples include
blocks of tissue (4 cm3) or several millilitres of fluid
placed in a sterile closed container. Anaerobic transport
medium is essential for swabs.
In suspected enterotoxaemia cases, where the demon-
stration of a specific toxin is required, at least 20 mL of
ileal contents should be submitted. A loop of ileum with
contents, tied off at each end, is acceptable or the ileal
contents drained into a secure sterile container.
Sample Submission
Laboratories usually supply a variety of sample containers
and transport media. The laboratory should supply sample
submission forms. These forms must be completed by the
veterinary practitioner and must give specific details of the
tests required as well as clinical history, differential diag-
noses, vaccination history, therapy, age of animal, etc. The
latter will enable the microbiologist to choose the most
appropriate tests and to avoid unnecessary expenditure. A
complete history will also allow the laboratory to identify
a particularly urgent situation and expedite the processing
of the sample. In certain instances owners may be con-
cerned that information in relation to their animals is kept
confidential. In such circumstances the clinician can assign
a code name or number to the animal and the owner but
should never omit clinical detail that will facilitate the
selection of the appropriate tests and the interpretation of
the results. The laboratory must be notified if the differen-
tial diagnosis includes a fungal infection or any agent that
is potentially infectious for people.
6
Many laboratories only set up certain tests on particular
days or at a designated time each day. Clinicians need to
familiarize themselves with the laboratory timetable in
order for them to provide an efficient service to their own
clients. An awareness of the time it takes to perform certain
assays is essential. Some assays may take weeks to com-
plete. Where certification is required, samples should be
submitted in good time and allowance should be made
for repeat testing and/or the collection of a second sample
if necessary.
The clinician should contact the laboratory in advance
if they are not a regular client or if the tests required are
not routine. It is essential to give the laboratory adequate
time to prepare for the receipt of a sample which requires
specialist testing. It is inadvisable, for example, to submit
a sample that needs to be passaged on a cell line that the
laboratory does not use on a regular basis without prior
discussion. In such circumstances the sample may have to
be stored for days if not weeks, while cells are resuscitated
from the freezer. In certain cases it may be necessary to
forward the sample to a specialist laboratory. If samples
are being submitted to a laboratory in another country an
import licence may be required.
Samples should be collected and delivered to the labo-
ratory as early in the day as possible so that processing can
commence on the same day. If a result is required urgently
this must be indicated on the request form and the head
of the laboratory should be notified in advance of the
arrival of the sample. Prompt delivery to the laboratory
will maximize the possibility of obtaining a diagnosis.
Viruses only replicate in living cells and a delay in sample
submission may result in loss of viability. The transit time
needs to be minimized. Samples should be transported in
contact with cold packs or wet ice. If transportation to the
laboratory is delayed, most samples should be held in the
refrigerator at +4°C rather than frozen. Serum harvested
from clotted blood samples can be stored frozen for
extended periods.
The labelling of samples must be clear and unambigu-
ous. Samples must be submitted individually in separate
leak-proof containers that are clearly marked, indicating
the identity of the sample (tissue, exudate, etc.), animal
identification and the date of collection (Fig. 1.3). Con-
tainer caps should be screwed on tightly and taped, if
necessary, to avoid leakage. All samples sent in the post
must be labelled and packed in accordance with the regu-
lations of the postal authorities. Glass tubes and other
fragile containers must be adequately packaged to ensure
they are not broken in the mail. Samples must always be
surrounded by sufficient absorbent material to soak up the
entire sample in the case of breakage or leakage. In general,
triple packaging of diagnostic samples is required. The
secondary or outer packaging must be a rigid container.
The package should be clearly labelled with the words
‘biological substances’.
 Collection and submission of diagnostic specimens
Figure 1.3 Sturdy, leak-proof containers for transporting
microbiological specimens.
Interpretation of Diagnostic Results
There must be full co-operation between the laboratory
and the veterinary practice for the benefit of the patient
and owner. It is the responsibility of the clinician to collect
and submit the appropriate samples accompanied by spe-
cific requests or adequate history. It is the responsibility of
the microbiologist to interpret the results in relation to the
information supplied. The quality of the diagnostic activ-
ity undertaken by the laboratory depends to a large degree
on the clinical and epidemiological information supplied
by the veterinary clinician.
The following points are pertinent when interpreting
reports from a diagnostic microbiology laboratory:
• A negative diagnostic report does not necessarily
mean that the suspected microorganism is not the
aetiological agent of the condition. There may be
many reasons for the failure of the laboratory to
isolate and identify the pathogen, such as a
bacterium being overgrown by contaminants, a
virus or other fragile microorganism having died on
the way to the laboratory or the animal may have
stopped excreting the microorganisms before the
sample was taken. Unexpected negative results
should be discussed with the head of the laboratory
who may decide to perform additional tests for the
detection of the suspect agent. They may also be able
to advise on the collection of alternative samples or
suggest other possible causal agents.
• It must not always be assumed that the detection
of a virus or bacteria establishes a diagnosis. The
laboratory result must be interpreted in light of the
clinical signs, vaccinal history, etc. If the sample is
collected from a site normally colonized with
bacteria, including species that have the potential to
Chapter |1|
cause disease in certain circumstances, the culture
results need to be interpreted cautiously. Some latent
viruses may reactivate and bacteria proliferate in
response to a condition which they have not caused.
Apparently healthy animals can be subclinical
shedders of microorganisms such as salmonellae,
rotaviruses, enteroviruses or coronaviruses in faeces
and leptospires in urine. Even if these potential
pathogens are isolated and identified, the illness
or death might have been due to another cause.
• The clinician should always discuss unexpected
results with the laboratory. This may assist the head
of the laboratory in the early detection of a problem
with a particular technique. The microbiologist must
be prepared at all times to act on information
received and to critically appraise the procedures in
the laboratory.
• It is important that the difficulties associated with
some types of samples and the limitations of certain
assays are flagged to the clinician. This will help to
promote intelligent utilization of the laboratory and
to avoid unrealistic expectations.
• Bacteria such as members of the Enterobacteriaceae
are ubiquitous. Isolation of microorganisms may
represent contamination of the sample by faeces or
soil or their presence could be due to postmortem
invasion.
• Some bacteria such as salmonellae, leptospires or
Mycobacterium avium subsp. paratuberculosis may be
shed intermittently. A repeat sample following a
negative examination report might be worthwhile.
• Escherichia coli in diarrhoeal faecal samples from
farm animals is usually only significant if the animal
is under 10 days of age and if the E. coli isolate
possesses the K88, K99, F41 or 987P fimbrial
antigens, associated with enteropathogenicity. Pigs
are the exception as they are also susceptible to
colibacillosis soon after weaning and to oedema
disease in the growing period.
• Diagnosis of a mycotic disease thought to be due
to a ubiquitous fungus such as Aspergillus fumigatus
should always be confirmed histopathologically. It is
necessary to demonstrate the fungal hyphae actually
invading the tissue and causing a tissue reaction.
• Herd or flock vaccination programmes will modify
the interpretation of serological tests. Serological
diagnosis of recent viral exposure usually requires
the comparison of two samples collected
approximately two weeks apart.
• Conflicting results may reflect the sensitivity of
different assays. Thus a sample that is negative by
enzyme-linked immunosorbent assay (ELISA) and by
isolation may be positive by PCR.
7
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 Chapter
Bacterial pathogens: Microscopy, culture
and identification
MICROSCOPY
A good-quality microscope with a built-in light source
(bright-field microscopy) as well as low-power, high-dry
and oil-immersion objectives is required. A darkfield con-
denser is a useful addition necessary for the visualization
of unstained preparations such as those of spirochaetes.
Microscopes for microbiology require a higher degree of
resolution than those used for haematology or histopa-
thology. Figure 2.1 indicates the size of bacteria relative to
that of an erythrocyte and to viruses. Bacteria are measured
in micrometres (µm) which are 10−6 m while viruses are
measured in nanometres (nm) which are 10−9 m. Using an
oil-immersion objective lens on a light microsope, a mag-
nification of 1000 × can be achieved to visualize bacteria.
On account of their small size viruses are visualized by
using an electron microscope, which utilizes a beam
of electrons to achieve magnifications of the order of
100,000 ×.
Stained Smears from
Pathological Specimens
Stained smears made from lesions can yield a considerable
amount of information inexpensively and quickly. Table
2.1 summarizes the information that can be gained from
the various diagnostic staining techniques.
Preparing Bacterial Smears
Microscope slides are not always clean enough to use
directly from the supplier. Rubbing with a clean, soft cloth
© 2013 Elsevier Ltd
2
and passing quickly through the Bunsen flame may be
sufficient to remove a greasy film. If not, a mildly abrasive
liquid cleaner can be used followed by rinsing the slide
thoroughly and wiping it dry with a clean cloth.
A blunt scalpel and forceps should be kept in a con-
tainer of 70% ethyl alcohol. The instruments are flamed
and cooled before use. Afterwards they should be placed
into a container of disinfectant. When making a smear
from tissue lesions, the specimen is held firmly with
the forceps and the scalpel is used to scrape deep into the
material. A small amount of the scrapings is placed on
the cleaned microscope slide. Another clean slide is used
with a scissor action to prepare a thin smear. With liquid
or semiliquid specimens, a little of the sample is placed
on the slide with a sterile swab. The contents of the swab
are smeared over the surface of the slide, with the aim of
having thick and thin areas of specimen present. The
smears are allowed to dry thoroughly before proceeding
further.
Fixing the Smears
The reasons for fixing the smears include killing the veg-
etative bacteria, rendering them permeable to the stain
and ensuring that the material is firmly fixed to the slide.
Fixed and stained smears should be handled carefully as
not all bacteria, especially endospores, may have been
killed. After use, the stained smears should be autoclaved
or soaked in a reliable disinfectant (24–48 hours) before
discarding.
For routine staining the smears are fixed by passing the
slide, smear side up, quickly through the Bunsen flame
two or three times, taking care not to overheat the smear.
This can be tested on the back of the hand; the slide
should feel warm but not hot enough to burn.
9
Section | 1 | General Procedures in Microbiology

Light microscope Electron microscope

Poxvirus
10 µm 1 µm 100 nm
Staphylococcus
Adenovirus

Parvovirus

Brucella

Chlamydia
Red blood cell

Figure 2.1 Comparison of the relative sizes of a red blood cell, bacteria and viruses.

Table 2.1 Diagnostic uses of stained smears

Disease and species Specimen Pathogens Appearance in stained

affected smears

Gram stain
Abscesses or suppurative Pus or exudate Staphylococcus species Gram + cocci, often in clumps
conditions
Many animal species Streptococcus species Gram + cocci, usually in
chains
Trueperella pyogenes Gram + rods, pleomorphic
Corynebacterium Gram + rods
pseudotuberculosis
Pseudomonas aeruginosa Gram − rods
Pasteurella multocida Gram − rods
Fusobacterium Gram −, long, slender
necrophorum filaments, often staining
irregularly
Strangles Pus Streptococcus equi Gram + cocci, often in chains
Horses subspecies equi
Suppurative broncho- Pus Rhodococcus equi Gram + rods with tendency to
pneumonia or superficial coccal forms
abscesses
Foals and young horses

10
 Bacterial pathogens: Microscopy, culture and identification
Table 2.1 Diagnostic uses of stained smears—cont’d
Disease and species
affected
Dermatophilosis
(Streptothricosis)
Many animals, mainly in sheep,
cattle and horses
Canine nocardiosis and
canine actinomycosis
Bovine actinobacillosis
(wooden tongue)
Bovine actinomycosis
(lumpy jaw)
Clostridial enterotoxaemia
Sheep and calves mainly
Urinary tract infections
Dogs and cats
Dilute carbol fuchsin (simple) stain
Campylobacter infections
Infertility in cattle. Abortion in
sheep and cattle
Swine dysentery
Foot rot in sheep
Modified Ziehl–Neelsen (MZN) stain
Brucellosis in cattle, sheep,
pigs and dogs
Nocardiosis
Canine nocardiosis
Bovine mastitis
Chlamydial infections
Sheep and cattle: abortion
Lambs, calves and pigs:
polyarthritis
Cats: feline pneumonitis
Chapter |2|
Specimen Pathogens Appearance in stained
smears
Scabs Dermatophilus Gram +, filamentous and
congolensis branching with coccal
zoospores arranged two or
more across
Pus or thoracic aspirates Nocardia asteroides Both Gram +, filamentous
Actinomyces viscosus and branching, Nocardia spp.
MZN +
Granules from pus Actinobacillus lignieresii Gram − rods
Actinomyces bovis Gram +, filamentous and
branching
Scrapings from small Clostridium perfringens Gram +, fat rods. Large
intestine of recently numbers suggestive of
dead animal enterotoxaemia
Fresh, carefully collected Escherichia coli, Proteus Gram − rods
urine species, Staphylococcus Gram + cocci
species and Enterococcus Gram + cocci
faecalis
Vaginal mucus or foetal Campylobacter fetus Curved rods that can be in
stomach contents chains giving ‘seagull’ forms
Faeces or scrapings from Brachyspira Numerous long (7.0 µm) but
colon hyodysenteriae finely spiralled bacteria
Exudate from hoof Dichelobacter nodosus Rods with a knob on one or
both ends
Fusobacterium Long, slender filaments,
necrophorum staining irregularly
Foetal stomach contents, Brucella spp. Small, red coccobacilli in
vaginal discharge, clumps
placenta
Pus and aspirates Nocardia asteroides Long-branching filaments,
Sediment from many staining bright red
centrifuged milk
Small, red coccobacilli in
Cotyledons Chlamydophila abortus clumps. Similar to brucellae
Joint fluid Chlamydophila pecorum Small, red coccobacilli
Conjunctival scrapings Chlamydophila felis
Continued
11
Section | 1 | General Procedures in Microbiology
Table 2.1 Diagnostic uses of stained smears—cont’d
Disease and species
affected
Ziehl–Neelson (acid-fast-) stain
Tuberculosis
Cattle and other species
Poultry, other avian species and
cervical lymph nodes of pigs
Feline leprosy
Paratuberculosis (Johne’s
disease) in cattle and sheep
Giemsa stain
Anthrax in cattle, sheep
and pigs
Feline infectious anaemia
Avian spirochaetosis
Dermatophilosis
(Streptothricosis) Many
animals, mainly sheep, cattle
and horses
Polychrome methylene blue
Anthrax in cattle, sheep
and pigs
Wet preparations
Leptospirosis in many
animal species
Ringworm in many animal
species
Other suspected fungal
infections in many animal
species
Bovine trichomoniasis
12
Specimen
Suspect lesions
Suspect lesions
Scrapings from lesions
Faeces, smear from
ileocaecal valve area and
mesenteric lymph nodes
Blood smear from ear
vein or fluid from
peritoneal cavity (pigs)
Thin blood smear
Blood smear taken during
febrile period
Scabs
Thin blood smear from
ear or tail vein. Fluid from
peritoneal cavity (pigs)
Centrifuged deposit of
urine or kidney tissue
Hair and skin scrapings in
20% KOH
Tissue, exudates, biopsies
in 20% KOH
Purulent uterine discharge
in saline. Keep warm and
examine within one hour
of collection
Pathogens Appearance in stained
smears
Mycobacterium bovis Long, thin, bright red rods,
can appear beaded. Usually
not numerous in smears
Mycobacterium avium As above but larger numbers of
acid-fast rods usually present
Mycobacterium Large numbers of red-staining,
lepraemurium acid-fast rods present
Mycobacterium avium Fairly short, red, acid-fast rods
subspecies in clumps
paratuberculosis
Bacillus anthracis Purplish, square-ended rods in
short chains surrounded by a
reddish-mauve capsule
Mycoplasma haemofelis Small, dark-blue coccal forms
on red blood cells
Borrelia anserina Helical bacteria, 8–20 µm long
and 0.2–0.5 µm wide with
five to eight spirals
Dermatophilus Blue, filamentous and
congolensis branching with coccal
zoospores arranged two or
more across
Bacillus anthracis Blue, square-ended rods in
short chains surrounded by a
pinkish-red capsule
Leptospira serovars Helical, approx.15 µm long
and 0.15 µm wide. Appears
beaded with hooked ends.
Dark field microscopy
Microsporum and Chains of refractile, round
Trichophyton spp. arthrospores on hairs.
High-dry objective
Aspergillus fumigatus, Fungal mycelial elements or
Candida albicans and budding yeast cells. High-dry
others objective
Trichomonas foetus Protozoan agent with three
free flagella and an
undulating membrane. Low/
high-dry objectives
Another random document with
no related content on Scribd:
 of children—Increase of crime—Arrival of money from
England—Orphanage—Labour question—Kūmishah—
Village ruffian—His punishment—Prince’s accident—The
kalāat—Mode of bringing it—Invitation to the ceremony
—Procession—Gala dress of the prince—The arrival of
the firman—Assemblage of grandees—The kalāat—The
Kawam’s kalāat—Return to town—Sacrifice of an ox
CHAPTER XXIV.
I FALL INTO THE HANDS OF BRIGANDS.
A call to a patient—Start on post-horses—No horses—I carry
a lantern—The Bakhtiaris—Fall among thieves—They
strip me—And march me off—Mode of disguise of
thieves—Attacked by footmen—Division of spoils—Fate
of a priest—Valuing my kit—Ignorance of my captors—A
welcome sight—My escape—I get a horse—Reach
Yezdikhast—Old women get thorns out of my feet—Want
of hospitality of head-man of Yezdikhast—Arrive at
Kūmishah—Kindness of a postmaster—More robbers—
Avoid them—Am repaid for my lost kit—Fate of my
robbers 259
CHAPTER XXV.
SHIRAZ.
The Muschir—His policy and wealth—His struggle with the
king’s uncle—He is bastinadoed—His banishment to
Kerbela—The Kawam—Mirza Naim—Siege of Zinjan—
Cruelties to Mirza Naim—Reply to an author’s statement
—Cashmere shawls—Anecdote—Garden of Dilgoosha
—Warm spring—“Sau-Sau-Rac”—The Well of Death—
Execution—Wife-killing—Tomb of Rich—Tomb of Hafiz—
Tomb of Saadi—A moral tale—Omens—Incident at tomb
of Hafiz 270
CHAPTER XXVI.
SHIRAZ—PERSIAN CUSTOMS.
The Tazzia—Persian pulpit—Prince’s flirtations—Month of 279
 mourning—Details of performance—Breast-beaters—
Hymn in honour of the king—The performers—
Processions—Detail of the tragedy—Interludes—
Rosehkhaneh—The Ramazan—The fast—Hospitalities
—Zalābi—Religious affectation—Reading poetry—A
paraphrase—A quotation—Books and their covers—
Calamdans—Writing a letter—Sealing—Specimen of an
ordinary letter—Apparent piety—The evil eye—
Talismans—I procure one
CHAPTER XXVII.
SHIRAZ.
Bagh-i-Takht—Jews’ burial-ground—Christians’ cemetery—
Its desecration—Sergeant Collins’s murder—Capture
and execution of the robbers—How it was brought home
to them—Memorial to Collins—Health of the staff—
Persians as servants—Persian cuisine—Kabobs,
varieties of—English dinners—Confectionery—Fruits—
Vegetables—Pickles, etc.—Cook-shops—Trotters—
Mode of selling meat—Game—Eggs—Wild vegetables—
Potatoes—Disinclination to use new seeds, and its
cause—Narcissus—General use of flower decoration—
Tame birds—Wild birds—White ants—Damaging the line
—Hamilton poles 292
CHAPTER XXVIII.
BEASTS, BIRDS, FRUITS, AND FLOWERS.
Tamed pigs—“Marjahn”—Mongoose—Persian cats—Their
value—Van cats—A fierce cat—How to obtain a Persian
cat—Grey-hounds—Toolahs—Watch-dogs—Monkeys—
Tame lions—Tame and cage birds—Superstition
concerning house-snakes—I kill a clockwinder—Wild ass
—Fighting rams—Tame partridges—Gardening—
Ordinary flowers—The broom-plant—Vine-culture—
Quinces and pomegranates—Orchards—Garden parties 302
CHAPTER XXIX.
 PERSIAN CHARACTER, COSTUMES, AND MANNERS.
Character of the Persians—Exaggeration—Mercifulness—
Anecdote—Costumes of men—Hair—Beards—Arms—
Costumes of women—Jewellery—Glass bangles—Nose-
rings—Painting of the face—Tattooing—Hair—Out-door
costume—Dress of children—Their manners—Strange
custom—Love of mothers—The uncle—Cousins—
Slaves—Servants—Slavery 314
CHAPTER XXX.
TRAVELLING—ART WORK—FOODS.
Travelling—Difficulties of posting—Saddles and bits—Cruel
joke—Old stories—Pastimes—Enamels—Persian
pictures—Curio buyer—Carvings—Metal-work—
Calligraphy—Kahtam—Incised work on iron—
Embroideries—Silver-work—Washing of linen—Ironing—
Needlework—The bath—Washing the hair with clay—
Bread and baking—Unleavened bread—Other kinds—
Travellers’ food—Inordinate appetites—Food of the poor 328
CHAPTER XXXI.
EDUCATION—LEAVE, AND RETURN VIÂ INDIA.
Education—Schools—Punishments—Love of poetry—
Colleges—Education of women—Religion—March to
Bushire—Extremes of cold and heat—Good luck—Go
home to England—Leave viâ India—The “Boys”—Lisbon
—Algiers—Port Said and Suez—Jeddah—Donkeys—
Coral reef—Sea-slugs—Aden—Madagascar oranges
—“Grimes”—Kurrachee—Drives—Visit to the alligators
at Muggerpir—Disgusting scene—A legatee—Black-
wood furniture—A lost bargain—Persian Gulf—Bushire
—Leave for Shiraz 337
CHAPTER XXXII.
FROM THE PERSIAN GULF TO ISPAHAN.
Our start for Shiraz—Camp out—Borasjūn—Spring at Dalliké 347
 —Kotuls—Kazerūn—Buy a horse—A tough climb—
Place of Collins’s murder—Arrive in Shiraz—Hire a
house—Settle down—Breaking horses—Night marching
—Difficulties of start—Mūrghab—Find our muleteer and
loads—Abadeh—Yezdikhast—Kūmishah—Mayar—Marg
—Arrive in Julfa
CHAPTER XXXIII.
JULFA.
Hire a house—Coolness of streets—Idleness of men—
Industry of women—Stone mortars—Arrack—Hire a
vineyard—A wily Armenian—Treasure-trove—The
“Shaking Minarets”—A hereditary functionary—A
permanent miracle—Its probable explanation—
Vaccination—Julfa priests—Arrack as an anæsthetic—
Road-making—Crops of firewood—Fire temple—Huge
trees—The racecourse—Disappearance of ancient brick
buildings—Donkeys—Healthiness of Julfa—Zil-es-Sultan
—His armoury—Prospects of the succession to the
throne—Bull-terriers—Mastiffs—Politeness and
rudeness of the prince 359
CHAPTER XXXIV.
JOURNEY TO AND FROM TEHERAN.
Proceed to Teheran—Takhtrowan—Duties—Gulhaek—Lawn- 368
tennis—Guebre gardener—A good road—The Shah—
Custom of the Kūrūk—M. Gersteiger—Cossack
regiments—Austrian officers—New coinage—Count
Monteforte—New police—Boulevard des Ambassadeurs
—English Embassy—Tile gates—Summer palaces—
Bazaars—Russian goods—Demarvend—Drive to
Ispahan—Difficulties of the journey—Accidents—Danger
of sunstroke—Turkeys—Keeping peacocks—Armenian
tribute of poultry—Burmese and Japanese embassies—
Entertainment and fireworks—Cruel treatment of Jews—
Oil paintings—Bahram and his queen—Practice makes
 perfect—Pharaoh and the Red Sea—Pharaoh and the
magicians
CHAPTER XXXV.
WE RETURN VIÂ THE CASPIAN.
New Year’s presents—Shiraz custom—Our cook’s
weaknesses—He takes the pledge—And becomes an
opium-eater—Decide to go home—Dispose of kit—Start
for Europe—Our own arrangements—Diary of our
journey home—Arrival 379
APPENDIX A.
Table of Post Stages and Ordinary Marches from Bushire,
Persian Gulf, to Teheran 410
APPENDIX B.
Duration of our Journey from Ispahan to London 412
APPENDIX C.
Travelling in Persia 413
APPENDIX D.
RUSSIAN GOODS VERSUS ENGLISH.
The Karūn River route—The best means of reaching the
Commercial Centres of Persia—Opinions of Experts—
Wishes of Merchants 417
Glossary of Persian Words 420
Index 429
LIST OF FULL-PAGE ILLUSTRATIONS.
SOLOMON IN ALL HIS GLORY Frontispiece.
PERSIAN BAND To face page 91
FEMALE DANCERS AND EQUILIBRIST ” 114
THE TOMBS OF THE KINGS (NAKSH-I-
RŪSTAM) ” 119
ARMENIAN WOMEN ” 132
THE BASTINADO ” 147
THREE SHOPS IN BAZAAR ” 189
THE PUL-I-KOJŪ ” 195
TENT LIFE—WANDERING TRIBES ” 262
A ROSEH-KHANA, OR PRAYER-MEETING ” 283
LION AND LUHLIS ” 307
MIDDLE-CLASS PERSIANS—PERSIAN BOY ” 317
OUTDOOR DRESS OF PERSIAN WOMEN ” 325
DR. WILLS’S HOUSE IN JULFA ” 360
A CHUPPER-KHANA, OR POST-HOUSE ” 386
 IN THE
LAND OF THE LION AND SUN.
 CHAPTER I.
I GO TO PERSIA.

Wanted, a doctor—The Director-in-chief—Doubt and distrust—Simple advice—Am

referred to ‘Hadji Baba’—My kit—Saddle for riding post—Vienna—Rustchuk—
Quarantine—Galatz—Kustendji—Constantinople—Turkish ladies—Stamboul
—I have my hair cut—“Karagews”—Turkish coffee—A philo-Turk—Shooting
party—The theatres—The Opera—Armenian theatre—Gambling-house—A
Bashi-bazouk—We leave, viâ the Black Sea—The Russian captain—
Unarmed vessels—White Crimean wine—Foreign wines in Russia—Deck
passengers—Sinope—Batoum—Poti—The post-house—Difficulty in getting
food—Travelling en tröika—Kutais—A tarantass—Apply for horses—An
itching palm—We start—Tiflis—Lecoq’s beer—A happy reprieve—The joys of
travel—Chief of the Telegraph in Tiflis—Uniforms—Persian Consulate—
Coffee and pipes—Smoking an art—Effects on the tyro—Tea—The Consul—
His age—Dyeing the hair—The Opera, varied costumes at—The Tiflis ballet—
Leave Tiflis—Erivan—The Pass—We lighten our load—Hotel—Washing—
Nakchewan—Julfa, the frontier of Persia.

I think that there is no more painful position than that of the young
medical man. I had “passed,” and had got my qualifications. An
assistant I did not wish to be, and I therefore consulted the
advertisement columns of the Lancet, and was prepared to go
anywhere, if I might see the world, and have what Americans call a
good time.
At my first attempt I came on an advertisement of three
appointments, under the Indian Government, in Persia; the address
was the Adelphi. Off I started for the Adelphi, which I had always
looked on as a neighbourhood full of mystery, and whose inhabitants
were to be mistrusted. Timeo Danaos.
A first-floor—this looked well. I knocked, was told to enter. Two
gentlemen, kneeling on the floor, looked at me in a disturbed
manner. The whole room is strewn with sheets of written foolscap,
and it appears that I have arrived inopportunely, as official
documents are being sorted. I am asked to take a seat, having
stated to the elder of the two that I am come to see the director on
business.
Now I couldn’t at that time fancy a director who knelt—in fact, my
only idea of one was the typical director of the novel, a stout,
bechained man—and my astonishment was great at being quietly
informed by one of the gentlemen that he was Colonel G⸺, and
should be glad to hear anything I had to communicate. I stated my
wish to obtain such an appointment as was advertised; the duties,
pay, &c., were pointed out, and I came to the conclusion that it would
suit me as a “pastime” till the happy day when I should have a brass
plate of my own. But if my ideas of a director were lofty, my ideas of
a colonel were loftier; and I said to myself, one who combines these
two functions, and can be polite to a humble doctor—must be an
impostor.
I was asked for my credentials. I gave them, and was told to call in
the morning, but distrust had taken hold of me; I got an ‘Army List,’
and, not finding my chief-that-was-to-be’s name in it, I, forgetting that
we had then an Indian as well as an English army, came to the
conclusion that he must be an impostor, and that I should be asked
for a deposit in the morning, which was, I believed, the general way
of obtaining money from the unwary.
With, I fear, a certain amount of truculent defiance, I presented
myself at the appointed hour, and was told that my references were
satisfactory, that a contract would be drawn up that I should have to
sign, and that I should be ready to start in a fortnight; but, rather to
my astonishment, no mention was made of a deposit. “I think there is
nothing more,” said Colonel G⸺.
This, I concluded, indicated the termination of the interview; and,
after considerable humming and hawing, I came to the point, and
blurted out that, after searching the ‘Army List,’ I couldn’t find any
Colonel G⸺, and that no one had ever heard of the Telegraph
Department in Persia.
Instead of being annoyed, the Colonel merely asked if I knew any
one at the War Office. As it happened I did. “Well, go to him, and he
will tell you all about it.”
Off I went to the War Office, found my friend, and, to his horror,
told him that I wanted to know if the Persian Telegraph Department
existed or not, and if the director was or was not a myth. He easily
satisfied me, and I felt that I had been stupidly suspicious.
I then announced to my friends and relatives my probably
immediate departure for Persia. Strange to say, they declined to see
it in any other light than a peculiarly elaborate and stupid joke.
Instead of congratulations, I was treated as an unamiable and tiring
lunatic, and from none of my friends was I able to get any
information as to Persia. One man had a son in Baghdad! but it was
no good his writing him, as it took six months to get an answer.
After a day or two I again presented myself at the office, and I had
the country described to me, and various recommendations as to
outfit given me, and I also was introduced to Major C⸺, the
assistant-director. His advice was delightfully simple. “You’ll be able
to wear out all your old clothes; don’t buy any new ones; have a
‘Dayrell’ bridle; get nothing but flannel shirts.” Colonel G⸺ certainly
took great trouble to explain to me all about the country, and, taking
me out to lunch with him, bought me Morier’s ‘Hadji Baba,’ saying,
“When you read this you will know more of Persia and the Persians
than you will if you had lived there with your eyes open for twenty
years.” This is going a long way; it is seventeen years since I went to
Persia, and I read ‘Hadji Baba’ now, and still learn something new
from it. As Persia was in Morier’s time so it is now; and, though one
sees plenty of decay, there is very little change.
Two other candidates came forward, to whom I was deputed to
explain matters. They accepted the conditions, and, the deeds being
prepared, we all three went to the India Office and signed a contract
for three years.
On going to the Adelphi I was told that a sum of one hundred
pounds had been handed to each of my two colleagues to take them
to Persia. But I was glad to seize the opportunity kindly given me by
Colonel G⸺ of travelling with him, and he told me to meet him in
Vienna on a certain day.
 I had now no time to lose, and proceeded to buy my kit; what that
kit was it is as well the reader should know.
I got enough ordinary clothing for three years, such as we use in
England for morning or country wear, also two pairs of riding-boots;
these fitted me, and were consequently useless, for I soon found that
in riding long distances boots much too big are the thing, as then the
foot is neither cold in winter or crippled in summer; a knife, fork, and
spoon, to shut up; a revolver; a small bradawl, with the point buried
in a cork, for boring holes in straps; a military saddle (hussar
officer’s), with wallet-holsters and a high cantle (this cantle keeps
one’s rugs off one’s back when riding post, which is the only way of
quick travelling in the country); a double-barrelled fowling-piece
(nearly useless). My kit was packed in a couple of bullock-trunks,
and my saddle sewn up in my rugs, which were thick and good. I
also had a blanket-lined waterproof sheet.
I gave myself a week in Paris previous to my nominal start, and
thence I proceeded to Vienna, to be ready to leave with Colonel G
⸺ as soon as he arrived there.
I went to the “Golden Lamb,” a very comfortable hotel which the
Colonel had chosen, and beguiled my time pleasantly enough in
going nightly to the theatre to hear Offenbach’s operas done in
German. I saw ‘Bluebeard,’ ‘La Belle Hélène,’ &c. I was a fortnight in
Vienna, and I began to pick up a smattering, for, of course, the
German learnt at school is useless; my Offenbach system I found
more effectual than the usual one of “the gardener’s wife has
brought the hat of the merchant’s little boy,” &c.
A week after the Colonel’s arrival our stay in Vienna ended. We
left for Basiatch (by rail twenty-seven hours); slept there, and started
early in the morning for Rustchuk by steamer. There we found that
passengers from up the river were in quarantine; and the letters
were taken with a pair of tongs, with immense precautions, for
fumigation; we were advised not to land, as we should certainly have
to go to the lazaretto; and we were told that if we quietly went on to
Galatz, and said nothing, we could return the next day as from a
healthy port.
 We were lucky in taking the advice, as a passenger did venture on
to the lighter, and was, willy-nilly, marched off to what we learnt
afterwards was a six weeks’ quarantine.
We went on to Galatz, which we reached the next day.
Galatz is like a rural Wapping, but muddier. We went to bed, to find
ourselves under weigh in the morning. We soon got to Tchernavoda,
which seemed a mere village. There we landed, and thence, by a
very slow train indeed, to Kustendji. At this place we heard the
ravages of the cholera had been very great. We slept there that
night, and started at noon next day for Constantinople by steamer.
It blew hard, and we were very glad indeed to find ourselves in the
Bosphorus. There the scenery became splendid; no description of
mine can do justice to the castles and palaces hanging on the
water’s edge; the crowded picturesque villages that were reflected in
the clear blue water; the shoals of porpoises that accompanied the
ship at full speed, ploughing the water with a loud noise, and then, in
their course, leaping, still continuing the race, from the water; and
then entering it again amid a shower of spray. This wonderful scene
continued for eighteen miles. At 5 p.m. we anchored in the Golden
Horn. The scene was indescribable; all I had ever seen or read of
paled before it. We were too late to land, as one cannot do so after
sunset.
Next morning we went ashore in a caique, rowed by very
picturesque boatmen in white kilts, passed the Custom House, and
went straight to Misseri’s, preceded by our baggage, borne by three
porters. These “hammals” bear gigantic burdens, and as in most
Eastern towns there are no carriage-roads, they are of great use,
and generally form a distinct corporation.
At Misseri’s the Colonel was well known, having stayed there
several times before. In Constantinople, happily for me, instead of
going on at once, my chief was delayed by orders from home for
nearly two months; and I was enabled to see a good deal of the
town.
 Great was my delight to watch the Turkish ladies, their muslin
yashmaks lending a fictitious delicacy to their complexions, going
about in handsome carriages. Innumerable were the mysterious
stories I heard after table d’hôte of these veiled beauties. Many a
time have I gone on long expeditions into Stamboul with Mr. Ayrton,
a brother of “Board of Works Ayrton,” who, with a thorough
knowledge of Turkey and the Turk, took me under his wing in his
daily pilgrimages to the most unsavoury but interesting nooks of the
Mahommedan portion of the city. We went to coffee-houses, and
listened to story-tellers; we dined on savoury kabobs; and, alas! I
well remember my philo-Turk friend persuaded me to have my hair
cut by a Turkish barber. It was only too well done; when the satisfied
shaver handed me the glass I was as a sheep before the shearer,
dumb, but with horror; my head was pink, so closely was it cropped,
and my only consolation was the remark of my introducer to oriental
life, that “in the East they generally did things thoroughly.”
I saw too the Turkish Punch (“Karagews”), a most immoral puppet;
and the mildest and most favourable description of him was that “his
manners were none, his customs disgusting,” but then my mentor
said he was “very oriental”—perhaps the terms mean much the
same thing.
As the coffee seemed particularly delicious in the native cafés, I,
after some trouble, ascertained the real receipt for coffee à la Turca
(not à la Turque), as they call it. Here it is; for each tiny cup (about a
small wineglassful), a teaspoonful of coffee fresh roasted, and
ground at once while hot to a fine powder in a brass hand-mill, or at
times pounded in a mortar, is thrown into a small and heated
saucepan; add the required quantity of boiling water. Place on the
embers; when it threatens to boil over, remove; replace, and remove
a second and a third time; serve. All the dregs go to the bottom. No
sugar or milk used—never clean the saucepan!
At these cafés long chibouques with yellow clay heads are
smoked, the heads being rested on a brass tray. A ball of live
charcoal is placed on the long-cut Samsoon tobacco (or if the
customer be liberal, Macedonian), the stem is jasmine or cherry
wood, and the grander the pipe the longer the stem; rich customers
bring their own mouth-pieces, which have a long inner conical tube
that fits any stem. These mouth-pieces are of amber, and are
frequently ornamented with a hoop of brilliants. The pieces of amber
are two in number, and if of large size and of good colour cost two
pounds, upwards to even five-and-twenty: the ordinary fashion is to
separate these two pieces by a thin circle of lapis-lazuli or other
stone.
The narghilé is also much used. It will be fully described as the
“kalian” further on. In it is smoked the tumbaku of Persia. A few
pence is charged for the whole entertainment of coffee, smoke,
shelter, and music, such as it is, generally a guitar or flute-player,
who is glad to play to order for a cup of coffee. The customers sit on
little low stools like the French church chairs without their backs. In
some of the grander cafés divans, and even chairs, are provided.
Mr. Ayrton had spent many years in Egypt. He wore a coat made
by a Turkish tailor, a shawl waistcoat and a fez, and with his cropped
grey hairs (it was his barber who operated on me) and his big
chibouque with the amber mouth-piece (he had a large collection of
them) with the ring of diamonds, he looked a thorough Turk, and I
fancy posed and was treated as such. I remember myself thinking
that the get-up was assumed for the purpose of getting a deeper
insight into Turkish life. From what I know now, I merely suppose
that, from his wearing the fez, he was, or had been, in Turkish
employ; all government servants in Turkey have to wear it. Dr.
Millengen, in whose arms Byron died, and who was an old
government employé (physician to three Sultans), wore it; so did his
son, who was in the Turkish Government Telegraph; and another son
of his, I afterwards met in the Turkish Quarantine Service at Teheran,
told me he wore it always while in Turkey.
I was introduced to a M. la Fontaine, a most enthusiastic
sportsman, and his many nephews, and by him I was given a day’s
cock-shooting, and there was plenty of it. As for me, I was an utter
muff and cockney, or rather town-reared; but had I not a new pin-fire
breechloader, and was it not my first day’s real shooting? And as I
really did shoot two brace, I returned a delighted but tired youth. That
night will be ever memorable. I ate my first pillaw, with fowls boiled to
rags in it, and followed by curds with thick cream on the top called
“yaourt.” How we all ate!
We had come from Pera, crossing in a steamer, and had to ride
some twenty miles on rough little ponies to the sleeping place, and—
horror of horrors!—on Turkish saddles. Now to the timid rider a
Turkish saddle is at first a delight, for to leave it without great effort is
impossible, and there is a pommel which is so high that it appears
the height of folly not to cling to it; but when one’s knees are in one’s
mouth, when one’s saddle is hard as iron and cuts like a knife, when
one has new and heavy shooting-boots on, and one’s
unmentionables have a tendency to ruck, besides having the glory of
carrying a forty-guinea gun slung (oh, demon cockney gun-maker!)
by a sling that slips along the barrel, and was highly recommended,
with the addition of one hundred loaded cartridges distributed over
the many pockets of a very new shooting-coat, in the sun, with a fur
cap on—is it to be wondered at that the sufferings of the tortured
Indian at the stake were child’s-play to what I endured without a
groan, and repeating constantly assurances of my delight and
enjoyment?—and remember, reader, we went at a brisk canter all
the time.
How glad I was to lie down! How grieved I was, at 4.30 a.m. the
next day, to be called, and, after a hurried wash, to start in the half
dawn in my tight and heavy boots! But the firing began; I forgot the
tightness of my boots, the stiffness of my back. Do you remember
how stiff you felt after your first riding lesson, my friend? and you
hadn’t one hundred loaded cartridges about you, and an intermittent
garotte with your knees in your mouth; and I thanked Heaven I need
not sit down, for weighty reasons.
Of course I fired wildly; of course I missed continually, but it was
my first day, and I never enjoyed anything so much in my life. I
hobbled bravely on till there was no more daylight, but I did feel
thoroughly done on getting in, and I did not enjoy my ride back the
next day.
I used to try and learn Persian in my idle hours, and I soon
mastered the printed character and could read fluently, but without
the slightest idea of meaning. Kind Colonel G⸺ gave me many a
lesson, but I fear that loafing in Stamboul by day and going to the
French or Italian theatre in the evening had greater attractions.
I was always passionately fond of the stage, and, as we were
always going in a day or two, I used, on the principle that I might
never be able to go to the play again, to go every evening.
Of course there was only a third-rate French company, but how
very good they were! The term “stick,” so justly applied to many of
our actors, could not be attached to any player in the little band. All
were good, and all were good all round, and though the leading man
might be everything in the drama, yet he didn’t object to play the
lover in the little vaudeville, and played it well. An Englishman, in the
event of anything so dreadful happening to him, would soon let his
audience see that he was only doing it under protest.
At the Opera the prima donna was ridiculously fat, and to a man
unmusical this somewhat destroys the illusion—but then the fauteuils
d’orchestre only cost ten francs. I also went to an Armenian theatre,
but it had the national characteristics, squalor and misery, and I did
not repeat the visit. I failed even to see an Armenian piece (if such a
thing exists), but sat out a fearful edition of ‘The Chiffonier of Paris;’
and I was told that all the pieces played in Constantinople (Pera) in
Armenian were mere translations.
Even the delights of gaming were permitted in Pera. A few doors
from Messeri’s was the Café “Flam,” as it was affectionately called
by the Pera youth. “Café Flamand” was, I fancy, its real title. Here
were played “pharaon” and roulette. I was recommended the former
game, for economical reasons—it took longer to lose a napoleon.
Nobody seemed to win at either game, but pharaon certainly “took
longer.” I was not tempted to make frequent visits, as I had played
for some small sums at Baden-Baden a year or two before. There
one was at least cheated fairly; here the robbery was open.
A few days after the New Year the Colonel told me that we should
really leave for Persia by the very first opportunity. I bid farewell to all
the kind friends I had made, had my photo taken in breeches, boots,
and revolver at Abdullah’s—a weakness every Englishman who
reaches Constantinople is guilty of. It does not do to be too oriental.
At Abdullah’s I purchased a fearful-looking type, marked a Bashi-
bazouk, and found it out afterwards to be the portrait of a man whose
acquaintance I made in Persia, the Dutch Consul in Bushire; but he
made a very good type, being a big man; and he literally bristled with
weapons, and seemed capable of any atrocities.
One fine afternoon, on January 5th, 1867, we were rowed on
board the Russian steamer Oleg. We had an English-speaking
captain, who was genial and communicative. My chief was confined
to his cabin; and as there was nothing to read and nothing to do, I
saw a good deal of the Russian. He told me that all the commanders
of their mail-boats were naval officers, and that all the mail-boats
could be turned into war-steamers at a few hours’ notice, merely
requiring the guns to be put into them: “so that, as you English don’t
let us have war-vessels on the Black Sea, we run a superior class of
mail-boat” (built, however, on the Clyde). And a very superior boat
she was.
I was told by the captain to avoid the high-priced wines, and stick
to white Crimean. This was a particularly delicious light wine, like a
good Sauterne; and I find, from after experience of Russian railway
buffets, which far exceed anything of the sort we have in grandeur,
that, as a rule, the liquor is simply fair red and white country wine,
the only difference being in price and label.
In some of these labels the Muscovite imagination fairly runs riot.
You see “Château d’Yquem,” “Schloss Johannisberg,” &c., but
nobody ever seems to drink them, and they are mere table
ornaments. The rich drink nothing but champagne of known and
expensive brands, and bottled stout; while the middle classes stick to
“piver” (Russian beer) and vodki.
Tea, in tumblers, was continually being served, with a big slice of
lemon in it. The deck passengers, among whom were many rough
Circassians, all armed to the teeth, cuddled down into the nooks of
the cargo, and managed to keep themselves warm as best they
could. They too always were drinking tea, but they adopted a plan to
economise sugar that I have noticed constantly among the Russian
poor: a bit of sugar is placed in the cheek, and then the tea is
swallowed in gulps; the poor fellows thus keeping up a sort of
delusion that they are swallowing sweet and hot tea, though the
mouth only, and not the tea, is really sweetened. There was none of
the exclusiveness of the Englishman. A made tea, and regaled B, C,
and D; then B treated the rest, and so on; when not asleep, eating,
or tea-drinking, the deck people were card-playing and smoking. The
short pipe was a good deal used, and passed from hand to hand,
while the trader class smoked the cigarette. All the men, and most of
the women, wore a sort of rough butcher-boot; and, from the state of
the roads at Poti, any other foot clothing for pedestrians would have
been impossible.
We lay to off Sinope on the 7th (here the Russians, our little
captain took care to remind me, destroyed the Turkish fleet), but
could not land passengers, a gale blowing. We changed steamers at
Batoum on the 10th.
The scenery at Batoum is very fine; the sea, without a wave, of a
deep blue; well-wooded hills and the Elburz range of the Caucasus
covered with snows forming the horizon. So warm was it here that
we lay on the beach throwing stones into the tranquil sea.
At last we arrived at Poti, being the fifth day from Constantinople.
We were put on a lighter with our baggage, and taken direct to the
Custom House; thence we got on a little steamer that was to take us
up the Riom river, and of this we had some twelve hours, the great
part of the time being occupied in getting aground, and getting off
again.
From Poti to Merand we went in a telega, en tröika, some sixty
versts, over what was rather a track than a road, in thirteen hours.
A telega, or road-waggon, is easily described as an oblong box on
wheels, and of the severest simplicity. The box is about five feet by
three feet six inches at the top, and five feet by three feet at the
bottom, with a plank in the front for the driver. There are no attempts
at springs; strength and lightness are all that is aimed at; these are
attained—also the maximum of discomfort. To this machine are
harnessed three horses: one trots in the shafts with a yoke four feet
high, the other two, in traces at either side, gallop. The harness is
rope, the driver often drunk.
Travelling thus is monotonous, and after a time very painful. To the
Russian officer, with his big pillow, little or no luggage, and plenty of
hay, a tröika is comparatively comfortable, for he can lie stretched
out, and be tolerably free from bruises, but, doing as we did, we
suffered grinding torments. One telega was full of our luggage, and
in the other we sat on a portmanteau of the Colonel’s; at each jolt we
were obliged to clutch the edge of the machine to prevent knocking
one against the other, and there was no support of any kind. To
people accustomed to ride on springs our sufferings would only be
apparent if they had once tried what it was to travel in this way for
many hours over the roughest roads, day and night and at full speed,
and without springs of any kind. When our hands got painfully
bruised we changed sides, and bruised the other ones, for we were
forced to hold on. When we were lucky enough to get a broadish
telega we got some hay, and sat on it, thus resting our knees.
On our way we only saw one woman and, say, a hundred men.
The country seemed to me very thinly populated after teeming
England. On our arrival at the post-house at Merand we were shown
a room with two plank bedsteads and a fireplace. I little thought that
in Persia the post-houses hadn’t even the plank bedsteads.
Neither of us could speak one word of the language; we tried
French, German, Italian, Turkish, Persian—all of no avail—and we
had no food. At last we obtained fire and a samovar, or Russian tea
urn; the first by pantomime, the second by looking fierce and
repeating the word.
We pointed to our mouths, heads were shaken (perhaps they
thought we wanted a dentist); at last I had a happy thought, and, by
drawing a hen and egg, and hopping about the room clucking, the
postmaster’s wife at last produced the required eggs; they then
brought bread and sausage, the latter much decomposed.
Colonel G⸺ was taken ill in the night, and I feared we could not
proceed. But by 8 a.m. (of the 12th) we were again on the road, and
did the thirty-four versts on a good military road by noon. The 12th is
with Russians New Year’s Day, and we found the town of Kutais for
the most part drunk and letting off its firearms.
Here our landlord informed us that there was an opportunity to buy
a tarantass, which we could dispose of when we reached the
Persian frontier or at Tiflis.
I was greatly delighted when the Colonel decided on purchasing
this very primitive carriage. Fancy an old-fashioned open carriage to
hold two, with cushions stuffed in prehistoric ages with hay, a
tarpaulin apron, a huge hood provided with a leather curtain which,
when dropped, plunged the traveller into black darkness, but kept
the wind and rain out; a gigantic box and boot, the whole slung on a
perch from four posts by thick straps, and having very small fore and
very large hind wheels, a plumb-line dropped from the top of the
latter being quite a foot beyond the bottom. But it kept us warm and
dry, would hold all the luggage, and would in theory enable us to
travel with three horses instead of six. We found out afterwards that
we had to take five, when we were lucky enough to get them.
I fancy the whole machine cost one hundred and fifty roubles, or,
at the then exchange, fifteen pounds. Then came a wheelwright, and
he took some seven hours at the wheels. At length, about five, all
was pronounced ready, and we sent our “padoroschna,” or permit to
take post-horses, to the postmaster for horses. Reply: “None just at
present; would send them over as soon as they came in.” To lose no
time, we carefully filled the boot with our luggage, and my bullock-
trunks were firmly roped on behind.
We took tea preparatory to our start, and laid in provisions of
bread, beer, &c., with a couple of fowls; for we were told we should
find nothing but black bread and hot water on the road. Still no
horses.
We went to the post-house, where we found nine beasts, but were
told that these were all reserved for special service. The Colonel
then smelt a rat; but what were we to do? the postmaster (a major)
was dining out, and no one knew where he was.