Bacteriological Status of Canine Milk and Septicaemia in Neonatal Puppies – a Retrospective Study

Results of the bacteriological examination of milk samples from 46 bitches were evaluated retrospectively, and correlated with findings of the bacteriological examination of organs from dead, septicaemic puppies in their litters (n = 33). The aim of this study was to investigate, in how many cases of clinical and subclinical mastitis of the bitches, the same bacteria can be detected in their septicaemic neonates. One group of lactating bitches was clinically healthy (group I, n = 38), whereas in eight bitches different puerperal disorders were found (group II). Twenty‐five septicaemic puppies were from group I, eight from group II. Out of a broad spectrum of bacteria isolated from the milk of clinically healthy and diseased bitches, only Escherichia coli, Klebsiella pneumoniae and/or ß‐haemolytic (haem.) Streptococcus sp. could be isolated from organs of their septicaemic puppies. This was the case in three bitches with mastitis and in one clinically healthy bitch only. Staphylococcus intermedius, although frequently isolated from canine milk, does not seem to be a cause of septicaemia in neonates. It is assumed that in most cases of neonate septicaemia, bacteria from the bitches’ milk are not the primary cause.     

Elevation of the bacteriological content of milk of clinically unaffected lactating bitches of a canine research stock

In the years 1987-1989 a bacteriological investigation of the milk was made in 44 clinically healthy postparturient bitches. In 30 animals bacteria most commonly isolated were staphylococci. 30.3% (40 samples) of the samples yielded pure cultures and 6.8% (9 samples) anacultures. Small numbers of bacteria were isolated in most of the samples; 67.4% showed moderate bacterial growth (less than or equal to 10(4)/ml sample). In vitro drug sensitivity tests revealed lincomycin, erythromycin and polymyxin B proved to be most efficient. Moreover neomycin, kanamycin and chloramphenicol showed low resistance rates while colistin sulfate and carbenicillin were less effective. A direct influence of lactiferous gland colonization with bacteria on mortality of puppies was not observed. With respect to clinical aspects, the application of antibiotics for sanitation of the lactiferous gland colonization with bacteria in clinically healthy bitches has to be considered carefully. However, for this the estimation of drug sensitivity tests is not satisfactory and should be completed by accurate bacteriological identification.     

Seriousness and stability of subclinical mastitis assessed by quarter milk serum albumin

Bovine Serum Albumin (BSA) concentration in quarter milk samples from 51 cows examined twice, at 1½ months intervals, was related to subclinical mastitis diagnoses and to the change in diagnoses from the first to the second examination. The BSA-concentration increased with increasing scores of the California Mastitis Test (CMT). The concentration of BSA was higher if bacteria were isolated as compared to negative bacteriological findings, and it was higher if “major pathogens” (MaP) (Staphylococcus aureus or Streptococcus dysgalactiae) rather than “minor pathogens” (MiP) (S. epidermidis or α-streptococci) were isolated. There was no interaction in the “effects” of CMT-score and bacteria on BSA-concentration. Quarters which were healthy and pathogenfree at first examination and had a non-specific mastitis at the subsequent examination, had significantly higher BSA-concentration at first examination than those which remained healthy. Quarters with non-specific mastitis at the first examination and infectious mastitis at the next examination had higher BSA-concentration at the first examinition than those which turned out with other diagnoses. Quarters with infectious mastitis and MaP at first examination which had a latent infection at the next testing, had a lower BSA-concentration than those with other second examination diagnosis. In general, BSA seems to be a more sensitive parameter than CMT for showing the early establishment of an inflammatory reaction in the alveolar tissue.     

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.     

Clinical and histopathological aspects of naturally occurring mastitis caused by Listeria monocytogenes in cattle and ewes

Listeria monocytogenes was isolated from the milk of two cows and two sheep with mastitis in one quarter and one udder half. The animals were observed over a period of 2-12 months. Clinical examination of the udder, bacteriological examinations and determination of somatic cell counts of milk samples were performed monthly. All four cases suffered from a subclinical mastitis characterized by an elevated somatic cell count (0.8-10.1 x 10(6) cells/ml), a persistent shedding of Listeria and by a normal appearance of the milk. The animals did not show any systemic reaction, but all animals developed an atrophy of the infected mammary gland. Histological examinations revealed a chronic interstitial mastitis with diffuse infiltration of lymphocytes, plasma cells and macrophages. All internal organs showed no abnormalities, no Listeria could be isolated. Listeria could however be isolated from the affected mammary parenchyma and from the mammary lymph node. The results of the bacteriological examination could be confirmed by means of PCR. Using PFGE, all the isolates from the same animal were identical. Immunohistochemical examination of the ovine mammary glands achieved a very strong immunoreactivity for CD5 cells. The mode of infection and the reaction of the immune system's defense of the ovine udders are discussed.     

奶牛急性乳房炎病原菌的分离与鉴定

[目的]为了掌握奶牛急性乳房炎致病菌的种类.[方法]通过采集病牛乳汁样品3份,分别对其病原茵进行分离与鉴定.检测[结果]表明:检出4种共13株细菌,其中致病性金黄色葡萄球菌6株;停乳链球菌5株;大肠埃希茵1株,乳房链球茵1株.[结论]该场奶牛的急性乳房炎是由致病性金黄色葡萄球菌、停乳链球菌、乳房链球菌和大肠杆菌共同引起...     

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.     

The prevalence of subclinical mastitis in dairy goats in Kenya

California mastitis test (CMT), direct leukocytes counts and bacteriological examination were performed on 630 milk samples from apparently healthy mammary glands of dairy goats comprising a mixed population of German Alpine, Toggenberg, Saanen and Galla crosses to find the prevalence of subclinical mastitis. The prevalence of subclinical mastitis was 9.8% according to CMT, 9.7% according to direct leukocyte counts and 28.7% by bacterial isolation during a 3-month period. The proportion of the bacteriologically positive milk samples was significantly (P <0.01) higher than that positive for CMT and direct leukocyte counts. There was a significant (P < 0.01) correlation between CMT and direct leukocyte counts. There was no significant direct relationship between bacterial isolation and CMT Bacterial organisms were isolated in 22.5% of the 568 CMT-negative milk samples. The results suggest that bacterial organisms isolated from the CMT-negative milksamples were either latent infections or did not stimulate any significant increase in somatic cell counts that could be detected by either the CMT or direct leukocyte counts. The observations of this study indicate that the mere presence of bacteria in goat's milk does not mean that the udder is infected and so does not warrant antibiotic therapy.     

Septicaemia in emerald monitors (Varanus prasinus Schlegel 1839) caused by Streptococcus agalactiae acquired from mice

The present study was performed to investigate both the identity and the source of the bacteria responsible for a fatal septicaemia observed in a group of three subadult emerald monitors (Varanus prasinus Schlegel 1839). The emerald monitors were necropsied and examined by light microscopy, including immunohistology, and by electron microscopy. Tissue samples were additionally submitted for bacteriological, virological and parasitological examinations. The virological and parasitological results were noncontributory, whereas the bacteriological investigation resulted in the isolation of gram-positive cocci which were characterized biochemically and serologically and by molecular analysis. The death of the emerald monitors was caused by a partially leukocyte-associated septicaemic infection with streptococci of serological group B of serotype V. Phenotypically and genotypically identical group B streptococci were isolated from the intestine of subadult mice, obtained from the feed used for the monitors. The genotypical characterization included an identical DNA fingerprint of strains of both origins, indicating the epidemiological relation between the feeding mice and the infections of the monitors.     

Limulus test--a rapid and simple method for the detection of endotoxins produced by gram-negative bacteria in mastitis milk

According to the present study the limulus amebocyte lysate test (LAL) seems to be a convenient test to detect endotoxin in milk from udder quarters with and without inflammation. The correlation between endotoxin concentration and the results from the bacteriological investigation of 79 milk samples was good (Table I). Determination of endotoxin in 20 milk samples from cases of acute clinical mastitis with high cell count and a negative bacteriological culture showed that all but one had an endotoxin concentration of greater than 1.0 ng/ml milk (Table II). By using a micromethod of the LAL it is possible to detect cases of mastitis caused by gram-negative bacteria about one hour after the sample has reached the laboratory. In a preliminary field study milk from 13 cases of acute clinical mastitis were tested by a modified limulus amebocyte lysate (LAL) test ("cowshed test"). A 100% correlation to bacteriological findings was observed (Table IV). By using the LAL test to detect mastitis cases caused by gram-negative bacteria great economic advantages and less risk for resistance problems can be achieved by using proper antibiotics. This is the fact in Sweden where the frequency of acute clinical mastitis caused by streptococci (100% of strains sensitive for penicillin) and Staphylococcus aureus (about 90% of strains sensitive for penicillin) is high (70-80%) and about 20% are caused by gram-negative bacteria, mostly E. coli.     

乳酸链球菌素治疗奶牛乳房炎效果观察

乳酸链球菌素（Nisin）是由乳酸链球菌产生的合有34个氨基酸的抗菌肽,对人体无害。本研究将患临床型乳房炎奶牛92头（107个乳区）和隐性乳房炎奶牛90头（90个乳区）随机分成试验组和对照组。试验组的治疗方法是在患病乳区灌注Nisin;临床型乳房炎以庆大霉素乳房内灌注作对照,隐性乳房炎的对照组不作任何处理。临床型乳房炎的Nisin治疗组细菌学治愈率和临床治愈率分别为60．8％和90．2％,而庆大霉素治疗组则分别为44.6％和91．2％,两者无统计学差异。隐性乳房炎经Nisin治疗后,高体细胞数奶牛的数量和牛奶中NAGase的活性均显著下降,细菌转阴率提高。乳酸菌发酵试验表明,经Nisin乳房内灌注治疗临床型乳房炎后,牛奶在第36小时恢复正常发酵,而庆大霉素治疗则需要72h。     

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.

     

The bacterial flora in the teat duct of ewes can protect against and can cause mastitis

We studied the possible effects of bacterial populations within the teat duct, in the pathogenesis of ovine mastitis. In experiment I, 32 ewes were allocated into group A (ewes from which we isolated (+++ growth) coagulase-negative staphylococci), B (ewes from whose duct we isolated (+ growth) coagulase-negative staphylococci) or C (ewes from which we isolated Bacillus spp.) and subdivided into A1, B1, C1 (n=4; challenged by deposition of 1.250 cfu of Mannheimia haemolytica into the teat duct) or A2, B2, C2 (n=4; used as uninoculated controls); group D (n=8) contained ewes with no bacteria in their teat ducts and were challenged as above. There were less bacteriological isolations of flora (P = 0.018) and challenge (P<0.05) organisms from A1 than from A2 and D ewes; the severity of pathological findings in A1 (summed up score: 27) ewes was smaller than in D (summed up score: 36) ewes (P = 0.038). No such findings were evident with B1 or C1 ewes (P>0.4). In experiment II, ewes (groups E and F, n=6) from whose duct we isolated coagulase-negative staphylococci (+ growth) were used; in group G (n=6) ewes with no bacteria in their teat ducts were included. Teat chapping was applied in E and G ewes. All E ewes developed acute clinical mastitis within 24 h after teat chapping, although we had carried out no challenge; there were more bacteriological isolations of flora organisms from E than from F and G ewes (P < 0.001); the severity of pathological findings in E (score: 28) was greater than in F (score: 3) or G (score: 14) ewes. In experiment III, eight ewes with no bacteria in their teat ducts were allocated into group H or I (n=4) and challenged into the teat (group H) or into the gland (group I) with 10(6) cfu of a Staphylococcus simulans recovered from the teat duct of a group E ewe. Group H ewes developed transiently clinical followed by subclinical mastitis (based on bacteriological and cytological evidence), whilst group I ewes developed severe clinical disease. We conclude that staphylococcal flora present in high numbers within the teat duct of ewes can afford some protection against invading microorganisms. However with impeded defence mechanisms of the teat, the same flora may invade the mammary parenchyma and cause clinical mastitis.     

Evaluation of a new non-hormonal substance for terminating pregnancy in bitches in clinical trials

A new non-hormonal contragestational agent (DL 717-IT), the 2-(4 chlorophenyl)-s-triazole |5,1-a|-isoquinoline, was tested in clinical trials to determine its efficacy for interrupting the pregnancy in the bitch. Five hundred and sixty-five bitches were given a single intramuscular injection of the drug between the first and 15th day after the mating. Of the 475 bitches that received 2–5 mg/kg, 430 (90-5 per cent) did not whelp at all, 15 (3-2 per cent) delivered a few stillborn or weak puppies that died some hours after birth, and 30 (6-3 per cent) delivered at least one normal live pup. Of 90 bitches treated with 3–5 mg/kg, 87 (96-7 per cent) did not whelp at all, two (2-2 per cent) delivered a few stillborn or weak puppies that died some hours after birth, and one (1 1 per cent) delivered normal live puppies. DL 717-IT, owing to the high effectiveness and minimal side effects, must be considered a very reliable drug for the interruption of pregnancies in the bitch.     

Efficacy and toxicity of tamoxifen citrate for prevention and termination of pregnancy in bitches

Five groups of bitches were given tamoxifen citrate (1 mg/kg of body weight) orally twice daily for 10 days. Drug administration commenced during late proestrus, day 4 of estrus, day 2 of diestrus, day 15 of diestrus, or day 30 of diestrus (n = 4/group). Nineteen of the bitches accepted natural mating by 1 or more of 3 stud dogs of known fertility (1 bitch did not). Twenty days after cessation of drug administration, ovarian, uterine, and hepatic specimens were obtained from each bitch in 4 of the groups. Pregnancy proceeded to natural termination in bitches of the remaining group (diestrous day 30). Pregnancy was not detected in any bitch of the proestrus, estrous, or early diestrous groups. Of 4 bitches of each of the remaining groups (diestrous day 15 and diestrous day 30), 2 aborted fetuses and/or resorbed placental remnants; the other 2 bitches in each of these groups had normal-appearing fetuses (diestrous day-15 group) or clinically normal pups (diestrous day-30 group). Of the 20 bitches given tamoxifen citrate, 5 developed endometritis with or without pyometra, and 4 of these had ovarian cysts. Although tamoxifen citrate is effective for preventing or terminating pregnancy in the bitch, the regimen used in the study reported here was associated with high frequency of pathologic changes in the reproductive tract.     

Composition and use of puppy milk replacers in German Shepherd puppies in the Netherlands

Enostosis or eosinophilic panosteitis is a common disease in young growing large-breed dogs, such as the German Shepherd, and the risk of developing the disease by 3-4 months of age is increased by a high calcium intake. The aim of the study was to investigate whether German Shepherd puppies raised on milk replacers receive more calcium and/or vitamin D than their requirements in the pre-weaning period and thus are at increased risk of developing skeletal diseases. To this end, we surveyed German Shepherd breeders in the Netherlands about the use of puppy milk replacers (PMR). The metabolizable energy, calcium, phosphorus and vitamin D content of the eight most used PMR were compared with that of bitch milk, as reported in the literature. The protein and fat content of most PMR were somewhat lower (range 24.4-33.2 g per 100 g on dmb and 18.3-37.5 g per 100 g on dmb respectively) compared with bitch milk (31.9 and 40.2 g on dmb respectively). The vitamin D content of one of the PMR samples was sevenfold the level recommended by the NRC (Nutrient Requirements of Dogs and Cats, National Academy Press, 2006) and threefold the average level of bitch milk. The clinical relevance of this high amount is questionable, as bitch milk contains mainly 25-hydroxy-vitamin D [3843 μg (96.1 IU) per 100 g on dmb] and only limited amounts of vitamin D [524 μg (13.3 IU) per 100 g on dmb], as was determined in this study. Dutch German Shepherd breeders tended to overfeed their puppies. We calculated that misguided use of PMR can increase the risk of excessive calcium, phosphorus and possibly vitamin D intake during a vulnerable period, potentially giving rise to bone and cartilage problems later in life.     

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.     

Milk substitutes and the hand rearing of orphan puppies and kittens

Growth in puppies and kittens during the suckling period is briefly reviewed. The nutrients required for this are discussed, as is their provision in bitch and queen milk. The use of milk substitutes in the rearing of orphan puppies and kittens is considered. The composition of a variety of milk substitute diets is determined, and the nutritive value of each product assessed.     

Use of clinical information to predict the characteristics of bacteria isolated from clinical cases of bovine mastitis

Farmers recorded the clinical signs of cows with clinical mastitis and submitted milk samples for bacteriological examination, so that the clinical signs could be correlated with the bacteriological findings. Odds ratios for the demeanour of the cow, the appearance of the milk, milk yield, udder texture, and the administration of parenteral antibiotics were calculated for mastitis cases classified in terms of their microbiology as either enterobacteriaceae, major Gram-positive pathogens, minor pathogens, 'no growths' or 'all other pathogens'. Animals infected with enterobacteriaceae had the highest odds of being reported as having a reduced milk yield, swollen or hard udders, watery milk and/or being systemically sick. A logistic regression model was used to predict the Gram-staining characteristics of the bacteria causing clinical mastitis. The clinical findings found to be significant predictors in the model were the demeanour of the cow and its milk yield. The regression model was used as a basis for a predictive test. Using a test data set, the sensitivity of the test was 28 per cent, its specificity was 96 per cent, the positive predictive value was 74 per cent and the negative predictive value was 80 per cent. The overall accuracy of these predictions was 79 per cent.