Long-term study of aerobic bacteria of the genital tract in breeding bitches.

The aerobic bacterial flora of the genital tract was characterized in 59 bitches in an 18-month study. The bitches represented 4 breeds and were from 3 kennels. Collection of vaginal swab specimens for bacterial culturing was performed every month, except during estrus when specimens were collected every week (n = 826). The capsule of the swab containing transport media was broken before specimen collection to moisten the tip, which helped to reduce the number of negative cultures. All bitches helped at least once during the study and, thus, had known reproductive functions. Pregnancy rates, litter sizes, and pup mortality were within normal limits. Pasteurella multocida, beta-hemolytic streptococci group G, and Escherichia coli were the most common bacteria isolated. Although these species generally were isolated from mixed cultures, pure cultures were obtained from 18% of the specimens. There was a tendency for the various breeds to differ in their vaginal bacterial flora. The flora also varied during the reproductive cycle. Pasteurella multocida was isolated significantly more often during proestrus, estrus, metestrus, and pregnancy, than during anestrus and the postpartum period, and beta-hemolytic streptococci were isolated significantly more often during proestrus than during estrus, pregnancy, or the postpartum period. Staphylococcus intermedius was almost exclusively found after parturition. Culture results were negative for only 5.2% of specimens cultured. On the basis of our findings, bacterial culturing of vaginal swab specimens from bitches without signs of genital disease is of little value.     

Significance of the vaginal bacterial flora in the bitch: a review.

The literature on the bacterial flora in the vagina of the bitch is reviewed. The bacterial flora in the vagina of healthy bitches is compared with the flora of infertile bitches and bitches with vaginitis, and the role of Brucella canis as a cause of infertility is evaluated. Several investigators have recommended that bitches with reproductive disorders be treated with antibacterial drugs, but there is very little knowledge of the effect of this therapy on the bacterial flora in the vagina.     

Some observations on the aerobic bacterial flora of the genital tract of the dog and bitch

The aerobic bacterial flora from the genital tracts of 143 bitches and 51 dogs was investigated. Beta haemolytic streptococci (group G), Staphylococcus aureus and Escherichia coli were among the organisms isolated from normal bitches in heat, abnormal bitches and normal dogs. Repeated sampling of two bitches during oestrus, and the dog to which they were mated, showed that bacteria were transmitted from bitch to dog, but did not become established. Guarded swab samples taken from the vagina collected less bacteria than samples from the vestibule. Numerous bacteria were seen in vaginal smears taken from bitches during oestrus, but numbers fell rapidly with the appearance of leucocytes in the smear at the onset of metoestrus.     

The pathological newborn in small animals: the neonate is not a small adult

The rate of perinatal death is variable but highest during parturition, immediately after birth and in the first days of life. Infectious diseases, above all bacterial, are the second most important cause of mortality after losses during parturition. A lot of factors are involved predisposing puppies and kittens to bacterial infections: respiratory distress, hypothermia, hypoglycaemia, dehydration, congenital abnormalities. E. coli, streptococci and staphylococci, Klebsiella sp., Pseudomonas sp., Enterobacter sp., Proteus sp. and anaerobes are regularly involved in bacterial infections in neonates. Postmortem findings especially document E. coli, Staphylococcus or Streptococcus species as causes of disease and death shortly after birth. The environment and mothers are suspected as sources of infection (vaginal discharge, milk, faeces, oropharynx, skin) for puppies and kittens. Genetic relatedness of bacterial strains in puppies and their mothers was found in staphylococci and E.coli. These results indicate that for repeated cases of bacterial infections in neonates diagnostic procedures of milk, vaginal and faecal swabs from bitches result in isolation of the responsible bacteria with a high probability and suggest that preterm treatment could help to control bacterial diseases and losses in pups.     

Aerobic Bacteria Occurring in the Vagina of Bitches with Reproductive Disorders

A retrospective survey was performed of aerobic bacterial species found in the vagina of 203 bitches with genital disorders, e.g. infertility, vaginitis, pyometra and puppy death. Escherichia coli, beta-hemolytic streptococci, Staphylococcus intermedius and Pasteurella multocida were the species most often isolated. From bitches with pyometra E. coli in pure culture was the most frequent isolate. In contrast, the majority of infertile bitches gave rise to mixed cultures, and no specific bacterial species was consistently associated with infertility. Thus, bacterial sampling from infertile bitches was concluded to be of low diagnostic value. Bacterial species isolated from the bitches having vaginitis were present in pure culture in 26.9% of the samples while nonspecific mixed cultures were obtained from 34.6% of the samples from these bitches. E. coli was the most frequently isolated bacterial species from bitches with dead puppies. However, in such cases it is important to relate the vaginal bacterial findings to autopsy findings and the results of bacteriological cultures of the pups.     

Bacterial infertility in camels (Camelus dromedarius): isolation of Campylobacter fetus

This study constitutes the first reported isolation of C. fetus subsp. venerealis from cervical swabs in breeding camels. 9 of 30 camels which were barren for a long period were found to be positive for C. fetus. The methods of isolation are described. Additionally, cervical flora of 30 camels were examined. 9 different bacterial species were isolated of which Staphylococcus aureus was most frequently found. All samples were negative for Trichomonas sp. and Streptococcus zooepidemicus.     

Colonization of neonatal puppies by staphylococci.

The composition of the normal staphylococcal flora of bitches and their litters held in a breeding unit was studied. The animals were sampled at a number of sites using moistened swabs. Six bitches were sampled daily, for 10 days, before whelping and then, together with four puppies per litter, at whelping (day 11) and at 1 and 7 days thereafter. Staphylococcus intermedius formed the predominant staphylococcal isolate. S. intermedius counts at the oral and nasal sites on the bitches did not change markedly before whelping and remained low (< 6 cfu/swab). Significant rises in the oral counts on both the bitches (P < 0.05) and puppies (P < 0.001) were then observed after whelping (days 11-18). Abdominal counts on both the bitches and puppies also rose (P < 0.001) after whelping. S. intermedius counts at the vaginal vestibulum of the pregnant bitches were found to be higher than at any other site sampled and did not alter markedly until whelping when a decrease (P = 0.05; days 10-12) was observed. S. intermedius was not found at the anal site in any of the six bitches and only transiently colonized five of the puppies.     

Fertility and whelping complications in bitches following correction of vaginal abnormalities

The fertility and whelping complications of normal bitches and of bitches that had had surgery to correct a vaginal abnormality of varying severities were compared retrospectively. Reproductive performance and whelping statistics were compared between 37 bitches that had been diagnosed with a vaginal abnormality and 37 age- and breed-matched bitches with no history of vaginal abnormalities. There were no significant differences in reproductive performance between the affected and control bitches. When analysed by the severity of the abnormality, pregnancy rates were significantly lower for the group of bitches with the most severe abnormalities compared with the group of bitches that had mild vaginal abnormalities. Furthermore, the bitches that had severe abnormalities were significantly more likely to require a caesarean section than those with mild abnormalities.     

Prediction of Parturition in Bitches Utilizing Continuous Vaginal Temperature Measurement

The objective of this study was to determine sensitivity and specificity of a body temperature decline in bitches to predict parturition. Temperature loggers were placed into the vaginal cavity of 16 pregnant bitches on day 56–61 after estimated ovulation or first mating. This measurement technique has been validated previously and enabled continuous sampling of body temperature. The temperature loggers were expelled from the vagina before delivery of the first pup. The computed values for specificity (77–92%) were higher than sensitivity (53–69%), indicating a more precise prognosis of parturition not occurring. In conclusion, our findings may assist interpreting vaginal temperature measurements in order to predict parturition in bitches.     

Escherichia coli strains isolated from the uterus and urinary bladder of bitches suffering from pyometra: comparison by restriction enzyme digestion and pulsed-field gel electrophoresis.

Pyometra (uterine inflammation with accumulation of pus in the uterus) is regarded as one of the most common illnesses in bitches. The ethiology and pathogenesis are complex with both hormonal and bacterial elements. The bacteria most frequently isolated from the uterine content is Escherichia coli.In this study, 84 E. coli strains from the uteri of 70 bitches suffering from the disease were examined and their DNA-profiles compared by restriction enzyme analysis and pulsed-field gel electrophoresis (PFGE). Through variations in DNA-profiles of the E. coli isolates, this study indicates that pyometra is caused by E. coli originating from the normal flora of the dogs and not by certain clones spread between animals.E. coli strains from the urinary bladder and the uterus of six of the bitches suffering from simultaneous urinary tract infection and pyometra were examined and compared as above. The DNA-profiles of the isolates from each of the six bitches were 100% identical. This study supports the theory suggesting that in cases of simultaneous urinary tract infection and E. coli pyometra, the urinary tract and uterus are infected with the same bacterial strain.To evaluate whether the uterus was infected with a single clone of E. coli or if multiple clones were present, eight to 16 colonies of E. coli isolated from pyometra samples from a further 10 bitches were examined. All bacterial colonies from the culture of the same bitch showed identical DNA-profiles.In 14 of the 70 bitches, two macroscopically different but biochemically identical E. coli colony types were isolated. The two colony types from the same bitch proved to have identical DNA-profiles in 13 cases and almost identical in the remaining bitch.     

An evaluation of positive contrast vaginourethrography as a diagnostic aid in the bitch

The contribution of vagino-urethrography to the diagnosis of disorders of the lower urogenital tract in 123 bitches was evaluated. It was found to be particularly valuable in animals with urinary incontinence, vaginal discharge, dysuria and physical vaginal abnormalities. Determination of vaginal volume in 69 bitches has enabled a dosage of contrast medium to be suggested.     

Use of exfoliative cytology for diagnosis of transmissible venereal tumour and controlling the recovery period in the bitch

Transmissible venereal tumour (TVT) is a rare vaginal tumour that can be treated surgically or cryosurgically as well as by radiotherapy or chemotherapy. Vincristine has been found to be very effective for treating TVT. Since vaginal secretion or discharge may contain neoplastic cells, the cytological identification of TVT cells is possible. The present study was carried out in 12 bitches. Vaginal smears were obtained with cotton swab from the anterior vagina and TVT suspected structures. The smears were stained according to Papanicolaou and assessed by light microscopy. Additionally the general condition of the patients was evaluated by haematological and radiographic examinations. In bitches with TVT vincristine sulphate was administered intravenously at weekly intervals. The total treatment period was three to six weeks until no atypical cells were found in the smear. This was the case after an average of 3.2 +/- 1.3 applications. Tumour masses became smaller and by this non-visible from the rima vulva after 4.2 +/- 0.7 applications. During the treatment, two of the 12 bitches (16.7%) suffered from vomiting and diarrhoea while three (25%) showed neutropenia. Twelve months after completion of treatment, the bitches were examined again and vaginal smears were taken in order to control the recovery process or a possible recurrence of TVT. No atypical cells were found in any vaginal smear. By this exfoliative cytology has proved to be a safe and easy method for TVT diagnosis and for observing the recovery process.     

Mastitis in Camelus dromedarius in Saudi Arabia

The correlation between camels' milk samples collected from abnormal inflamed udders and samples positive in the California Mastitis Test (CMT) was +0.803 (P less than 0.01). The bacterial count ranges of milk samples differed significantly (P less than 0.05) for those with a negative CMT and those with a positive CMT. Infection with many but not all bacterial species was associated with positive CMT results. The highest percentage of camel milk samples was included in the bacterial count range of 3.0 x 10(2) to 3.0 x 10(3) cfu/ml rather than in the greater than 3.0 x 10(3) cfu/ml range for most of the bacterial species. The most predominant bacterial isolates were Micrococcus spp., Staphylococcus aureus, Streptococcus spp. and Corynebacterium spp. followed by eight other flora. Chloramphenicol was the most effective antimicrobial agent of six tested against 118 bacterial isolates. Preliminary observations are made on chemotherapy of mastitis cases in camels.     

Diagnosis of Endometritis in the Bitch: A New Approach

There are a few investigations into endometritis in the bitch and its relationship with failure to conceive remains unclear. This may be because of the difficulty in collecting uterine samples for further investigations. Recently, transcervical catheterization by vaginal endoscopy has been introduced allowing the evaluation of the endometrium. In this study, uterine cytology and bacteriology were evaluated in 26 infertile bitches. Endometritis was bacterial in origin in most cases (70% of affected bitches), but these results may be underestimated, as some other pathogens (anaerobic bacteria, mycoplasms and fungi) were not investigated. Endometritis, in our opinion, should be investigated in each case of unexplained infertility in bitches. The method used here seems reliable although defining more accurate classification criteria will improve the efficiency of this non-invasive technique.     

Vaginal and uterine microflora of adult dogs

Aerobic and anaerobic microflora were identified and quantitated in 82 vaginal and 78 uterine samples obtained from mature bitches during different stages of the estrous cycle. The mean +/- SD of total bacterial counts/100 mg of vaginal contents of the 82 bitches was log 5.0 +/- 1.5, ranging from log 2.4 to log 8.8. The count at the estrous stage (log 7.8 +/- 0.7) was significantly higher (P less than 0.05) than that at the anestrus (log 4.4 +/- 1.0), pregnancy (log 5.9 +/- 1.3), and postpartum (log 5.1 +/- 1.5) stages. The common organisms isolated from the vaginas were Bacteroidaceae, streptococci, Pasteurella spp, and mycoplasmas. Organisms were isolated from 48 (68%) of 78 uterine samples. The range of total counts/100 mg of uterine contents was from log 1.6 to log 8.3. Staphylococci and mycoplasmas were frequently isolated from the uterine contents. Although many uterine microfloras were similar to vaginal microfloras, some uterine culture had a single isolate identified. There were no pathologic findings in most of the uteri. Seemingly, vaginal bacteria frequently flow into the uterus, yet they rarely cause uterine infection.     

圈养大熊猫食性转换前期肠道细菌多样性

为了探讨大熊猫食性转换前期肠道菌群的多样性与差异性。对3只处于夏季食性转换前期大熊猫粪便细菌总DNA进行ERIC—PCR分析,并构建16SrDNA克隆文库,采用限制性内切酶HinfI、MsPI对其进行RFLP（restric—tionfragmentlengthpolymorphism）及测序分析。研究表明,食性转换前期大熊猫肠道细菌主要由硬壁菌门和变形菌门组成。其中7月份样品中以梭菌属和醋杆菌属为主;8月份以八迭球菌属和埃希氏菌属为主;9月份以链球菌属和埃希氏菌属为主。此外在3个月中均发现一定比例的未培养细菌。结果表明：随着食性转换前期的过渡,大熊猫肠道菌群发生了改变。由硬壁菌门中的梭菌属渐变到以球菌属为主,由变形菌门的醋杆菌属渐变到以肠杆菌为主。     

Vaginal cytology and cervicovaginal mucus arborisation in the breeding management of bitches

Fifty bitches were mated on the basis of results of vaginal cytological and cervicovaginal mucus arborisation examination. When these were compared with a control group of 50 bitches mated on days 10 and 12 after the onset of proestrus there was a significant increase in pregnancy rate from 78 to 92 per cent. The litter size and bitch productivity was also increased, although this was not statistically significant. Forty-two per cent of bitches had two peaks of anuclear epithelial cells in the vaginal smear, separated by a mean of 3–6 ± 1–5 (SD) days. For these bitches, the maximum percentage of anuclear cells at the second peak was between 57 and 98 per cent, and the mean interval between the calculated day of the surge in plasma luteinising hormone (LH) and the second peak of anuclear cells was 4-4 ± 2-0 days. In bitches with one peak of anuclear epithelial cells, the maximum percentage of anuclear cells was between 43 and 94 per cent and the mean interval between the calculated day of the LH surge and the peak of anuclear cells was 1–9 ± 2-1 days. Arborisation of mucus collected from the anterior vagina was noted in all bitches; the mean interval between the calculated day of the LH surge and the peak of ferning was 2–3 ± 2-1 days. The collection of exfoliative cells by aspiration from the cranial vagina and the assessment of the anuclear cell index, may allow the identification of the fertile period after the surge of plasma LH. The anuclear cell index offers advantages over other methods of classification of exfoliative vaginal cells. When used in combination with examination of arborisation of cervicovaginal mucus this method can improve the breeding management of bitches.     

Uterine, cervical and vaginal microflora of the normal bitch throughout the reproductive cycle

Samples for microbiological culture were collected from the uterus of bitches using transcervical uterine cannulation (31 samples, 23 bitches) and from the uterus, cervix and vagina post mortem (19 bitches) at all stages of the reproductive cycle. Samples were cultured for aerobic and anaerobic bacteria and for aerobic mycoplasmas. Bacteria were always found in the uterus during prooestrus and oestrus (12 positive in 12 cultures) and rarely at other stages of the reproductive cycle: during anoestrus (one in 14) and other stages (none in 24). When microorganisms were detected at three sites post mortem, those found in the cervix and vagina were always of the same species as those found in the uterus. In six out of 13 instances, microorganisms were found in the cervix or vagina when none were found in the uterus. The mean number of isolates, number of bacteria seen in uterine cytology and bacterial growth were greater (P < 0–005) during oestrus and pro-oestrus than at other stages. Bacteria isolated from the uterus, in order of frequency, were Escherichia coli, Haemophilus species, α-haemo-lytic streptococci, Corynebacterium species, Streptococcus canis, Alcaligenes faecalis, Bac-teroides species, Pasteurella species and Proteus mirabilis. No mycoplasmas were cultured from the samples. This study indicates that the uterus of the normal bitch has a uterine microflora during pro-oestrus and oestrus that is similar to that of the vagina and cervix.     

Bacteriological findings in the canine uterus during Caesarean section performed due to dystocia and their correlation to puppy mortality at the time of parturition

Canine intrauterine bacteriological flora during dystocia is unknown. Thus, frequency (bacterial growth (not) detected), quality (species and number of different bacterial isolates) and quantity (colony‐forming units) of intrauterine bacteria in relation to in utero foetal death in 50 bitches undergoing emergency Caesarean section were investigated. Bacterial growth was quantified from single colonies, (+) (0.5), to strong growth, +++ (3) and was observed in 34 bitches (68%), with Staph. epidermidis (n = 12), Staph. intermedius‐group (n = 7), β‐haemolytic streptococci (n = 6), Staph. aureus, α‐ and γ‐haemolytic streptococci (n = 4 each) being most common and one to four bacteria per sample. Regarding the quantity, most often (n = 46) low growth was identified. In bitches with living pups only (group I), mean number of isolates was 0.78 ± 0.83 compared to 1.60 ± 1.10 (living + stillborn pups, group II) and 1.0 ± 1.15 (stillborn pups only, group III) and mean bacterial growth in groups I/II/III was + (1.0, quantity), + (1.4) and ++ (1.6). Taking just positive samples into consideration, mean number of bacterial isolates was significantly higher in group II compared to I (p = .0088). We concluded that the canine uterus cannot be considered free of bacteria during dystocia. Mean numbers of different bacterial isolates and quantity of bacterial growth are higher in bitches with in utero foetal death.     

Rapid immunohistochemical detection of Rhodococcus equi in impression smears from affected foals on postmortem examination

The first objective of this study was to develop an immunohistochemical procedure for rapid detection of Rhodococcus equi in impression smears from affected organs of foals on postmortem examination. The second aim was to demonstrate whether R. equi can be detected in smears of tracheal exudates collected from the same foals using an immunohistochemical method. Impression smears and cryostat and paraffin-embedded sections were made from the lungs and mediastinal lymph nodes of three foals (A, B and C) that had died of respiratory disease caused by R. equi, and also from the caudal mesenteric lymph node of foal A. Impression smears were made from the tracheal exudates of all foals. An affinity purified rabbit IgG was used for the immunohistochemical demonstration of R. equi. This antibody reacted with serotype 1 of R. equi in Ouchterlony's immunodiffusion and in the passive haemagglutination test, but not with other serotypes or with Streptococcus equi ssp. equi or Staphylococcus aureus, and failed to give an immunohistochemical reaction with Mycobacterium bovis or M. paratuberculosis. The immunohistochemical method proved to be of identical sensitivity to bacterial culture; moreover, from the lungs and mediastinal lymph nodes of one foal, R. equi could only be detected by this method. R. equi was demonstrated in smears of the tracheal exudates of all three foals. The results of this study indicate that the immunohistochemical method may be used for the rapid detection of R. equi in impression smears from the affected organs, especially abscesses, obtained postmortem, and possibly as a tool for diagnosing R. equi pneumonia in live foals by examining smears of tracheal aspirates.