Microbiologic evaluation of gallbladder bile of healthy dogs and dogs with iatrogenic hypercortisolism: a pilot study.

BACKGROUND: In people, hypercortisolism (HC) has been associated with acalculous cholecystitis and biliary dyskinesia, which may potentiate ascending biliary infections. In dogs, an association between HC and gallbladder disease recently has been documented, although the role of bacteria remains controversial. Furthermore, there is no information on the gallbladder bile microbial flora in healthy dogs. OBJECTIVES: To investigate the microbial flora in gallbladder bile in healthy dogs, the relationship between iatrogenic hyperadrenocorticism and bactibilia and possible changes in biliary microbial flora after cortisol withdrawal in dogs. ANIMALS: Six control dogs and 6 dogs treated with hydrocortisone. METHODS: Gallbladder bile obtained by percutaneous ultrasound-guided cholecystocentesis was cultured aerobically and anaerobically and examined cytologically before (d0), during (d28, d56, d84), and after (d28p, d56p, d84p) administration of hydrocortisone (8 mg/kg PO q12h). RESULTS: In the control group, 2/42 bile cultures yielded bacterial growth (Enterococcus sp.; Escherichia coli on d0) and 1/42 bile smears had cytological evidence of bacteria (d28). In the HC group, 2/42 bile cultures yielded bacterial growth (Enterococcus sp. on d28; Bacillus sp. on d28p) and 3/42 bile smears had cytological evidence of bacteria (d84, d84, d28p). All dogs remained healthy throughout the study period (168d). CONCLUSIONS AND CLINICAL IMPORTANCE: Based on the results of conventional bacterial culture techniques, gallbladder bile of healthy dogs periodically may harbor bacteria, which do not appear to be clinically relevant. A 3-month period of iatrogenic HC was not associated with bactibilia. A higher prevalence of bactibilia may be detected with micromolecular techniques.

[Field study of the prevalence and diagnosis of diarrhea-causing agents in the newborn calf in a Swiss veterinary practice area]. / Feldstudie zu Prävalenz und Diagnostik von Durchfallerregern beim neonaten Kalb im Einzugsgebiet einer schweizerischen Nutztierpraxis.

The prevalence of cryptosporidia, rotavirus, bovine coronavirus and Escherichia coli F5 (K99) in dairy calves with diarrhea and in healthy calves was established in a limited area served by a veterinary practice. Immuno-chromatographic rapid tests (FASTest Strips) were applied in the field and their results were compared to the ones obtained with standard methods (modified Ziehl-Neelsen stain, antigen-ELISA and cultivation). In 78% of the calves with diarrhea (n=46) and in 29% of the healthy calves (n=14), one or two agents were isolated. Of the diseased calves, 43% excreted cryptosporidia and in 46% rotavirus was isolated. Bovine corona virus and Escherichia coli F5 (K99) seemed to be of minor importance in the investigated population. Compared to the modified Ziehl-Neelsen stain or the antigen-ELISA, the FASTest Strips CRYPTO and ROTA were of very high diagnostic specificity of 100% each and their diagnostic sensitivity was 75% and 57%, respectively. Due to the low number of cases, the results of the FASTest Strips BCV and E.coli-K99 could not be interpreted. Although the diagnostic sensitivity of the FASTest Strips CRYPTO and ROTA--evaluated with standard methods--was not very high, their use in calves with acute diarrhea is recommended.

[Incidence of zoonoses in petting zoos and evaluation of hygiene measures to prevent the transmission to humans]. / Vorkommen von Zoonosen in Streichelzoos und Beurteilung der Hygienemassnahmen für die Ubertragung auf den Menschen.

In summer 2003, a study was performed in thirty Swiss petting zoos with the objective to determine the prevalence of zoonotic agents, and to describe hygiene measures implemented to reduce the risk of human infection. Fecal samples from different animal species were collected from the floor of pens to determine the prevalence of Salmonella spp., Campylobacter spp., verocytotoxin producing E. coli/ VTEC and Francisella tularensis. A questionnaire on hygiene measures, number of animals per species, housing system, care procedures and feeding was administered to every petting zoo to estimate exposure of visitors to zoonotic microorganisms. In total, 423 fecal samples were examined. Of these samples, 41 were positive for Campylobacter spp., which were mainly isolates from pigs and poultry (35% positive samples from each species). In pigs, 50% of the positive samples (6 samples) were typed as C. jejuni. The others were typed as C. coli (3) and C lan' (3), respectively. Five poultry isolates were typed as C. jejuni, and two as C. coli. Two samples were positive for Salmonella spp. Salmonella typhimurium was isolated from a goat, the other isolate could not be identified by serotyping. Neither Francisella tularensis nor verocytotoxin producing E. coli/ VTEC were found. The low prevalence of zoonotic microorganisms in Swiss petting zoos could be attributed to the cleanness of enclosures and animals, low stocking rates and good animal care. However, there is room for improvement concerning visitors' information on hygiene and hand washing. Furthermore, a strict separation between picnic - areas and animals should be enforced.

Experimental Chlamydia psittaci serotype 1 enteric infection in gnotobiotic piglets: histopathological, immunohistochemical and microbiological findings.

The enteric pathogenicity of the ovine C. psittaci serotype 1 isolate S26/3 was assessed using a litter of gnotobiotic piglets. In one group, eight piglets were inoculated at 3 days of age; at 10 days, two of these were re-inoculated. In a second group, six animals were mock-inoculated at 3 days of age as negative controls; subsequently, at 10 days, three of these piglets were inoculated with C. psittaci. The animals were observed for clinical signs, killed and necropsied sequentially between 4 and 17 days of age. At necropsy, specimens were collected for histopathology, immunohistochemistry and serology. Clinical manifestations consisted of sporadic slight softening of faeces observed between 8 and 12 days post inoculation (d.p.i.) in pigs inoculated at 3 days of age and between 4 and 6 d.p.i. in those inoculated at day 10. Histopathological changes were minimal and inconsistent and occurred almost exclusively in the small intestine in pigs of 15 days of age and older; they consisted of a slight shortening of villi, of a small number of tongue-shaped villi and of villous fusions. Immunohistochemistry revealed small numbers of chlamydial inclusions in the small intestinal enterocytes of only five pigs, all killed within 5 d.p.i. An ELISA run on faecal samples collected daily after inoculation from six of the pigs showed that chlamydial antigen was excreted in the faeces. In pigs inoculated at 3 days, chlamydial antigen was detected inconsistently before, and consistently after 9 d.p.i. Pigs inoculated at 10 days excreted antigen consistently after inoculation until the end of their observation period (8 d.p.i.). Infective chlamydiae were detected from the faeces of inoculated piglets using Vero cell cultures. Sera of all pigs were negative for anti-chlamydial antibodies using a complement fixation test. In conclusion, enteric pathogenicity of C. psittaci serotype 1 in a litter of gnotobiotic piglets proved minimal. The results, therefore, indicate that serotype 1 C. psittaci is not likely to cause enteric disease in conventionally reared pigs. Nevertheless, a potential role of swine in the epidemiology of this agent should be considered with regard to spread of Chlamydia to other species.

Elastase-producing microorganisms in horse lungs: their possible role in the pathogenesis of chronic pulmonary disease in the horse.

Seventeen out of 21 horses had pulmonary microbial organisms which reached considerable numbers in seven cases. Elastase-producing microorganisms from the environment (Streptomyces species and to a lesser extent Bacillus species) constituted 22 per cent to 99 per cent (mean 79 per cent) of the total growth. There was a considerable number of microorganisms with in vitro-produced elastases which were not or only slightly affected by horse serum. There was no correlation between numbers of organisms and pulmonary histopathological findings thus the significance of these microorganisms in the pathogenesis of alveolar emphysema is unknown. The growth of a strain of Streptomyces collinus/diastatochromogus isolated from the lungs was suppressed by fresh horse serum but not by decomplemented horse serum. Complement activation in response to this organism could contribute to airway inflammation through the production of mediators.

Streptomycin-resistant Escherichia coli as a marker of vulvovestibular contamination of endometrial culture swabs in the mare.

To investigate the vulvovestibular contamination of endometrial culture swabs in the mare, a liquid culture of a streptomycin-resistant strain of Escherichia coli was applied to the vulvovestibular area of mares and used as a marker of contamination of endometrial culture swabs. Prior to taking endometrial swabs, the perineal area was washed with soap, rinsed with water, and dried. Endometrial culture swabs were taken from mares that were in anestrus or diestrus and from mares that were in estrus. When a manual transvaginal swabbing technique was used, 22 of 24 endometrial swab specimens from 12 mares were contaminated with the experimental bacterial strain; culture of only one endometrial swab yielded more than nine colonies. When a speculum approach was employed, three of 12 swab specimens from 12 mares yielded between one and three colonies. The stage of cycle had no effect on the extent of contamination, but the proportion of positive cultures was significantly smaller when swabs were taken via a vaginal speculum approach, compared to a manual transvaginal approach. Complete preclusion of vulvovestibular contamination of endometrial swab specimens was not achieved; however, fewer than ten colonies can be expected even in mares in which the vulvovestibular area has been thoroughly contaminated with a broth culture, provided that the perineal area is adequately cleaned prior to swabbing.

Diagnostic investigation into the role of Chlamydiae in cases of increased rates of return to oestrus in pigs.

Cervical swabs and serum samples were taken from Swiss herds of sows with high rates of irregular return to oestrus (group A) and from control herds without reproductive problems (group B. The genital tracts of 21 slaughtered sows of group A were also examined. The swabs and genital tracts were screened for Chlamydiae by a new 16S rRNA PCR and the sera by an ELISA for Chlamydiaceae lipopolysaccharide. Chlamydophila (Cp) abortus was isolated from seven of the 65 swabs taken from group A but from none of the 128 swabs taken from group B. Chlamydia suis was present in swabs from both groups A (1.5 per cent) and B (2.3 per cent). In addition, Cp abortus was detected in 33.3 per cent of the genital tracts. Of the 193 sera tested, 61.7 per cent were positive, with no significant difference between group A (52.3 per cent) and group B (66.4 per cent). Chlamydia-like organisms were detected in 28.2 per cent of the swabs from group A and in 22 per cent of those from group B.

Demonstration of heterogeneous genotypes of Taylorella equigenitalis isolated from horses in six European countries by pulsed-field gel electrophoresis.

Forty-six isolates of Taylorella equigenitalis were analysed by pulsed-field gel electrophoresis (PFGE) after separate digestion of the genomic DNA with ApaI and with NotI. The isolates had been obtained from horses in six European countries and were classified into 18 genotypes. In Belgium, 2 genotypes were detected in 2 isolates, in England 9 among 15, in Finland 2 in 2, in France 2 among 10, in Sweden 3 among 5, and in Switzerland 3 among 12. Two English isolates and 4 French isolates gave identical PFGE profiles to those of Kentucky 188 from the United States. A common genotype was found in 5 isolates from Belgium and England and also in 10 isolates from France and Switzerland. The analysis of genomic DNA from 12 isolates of T equigenitalis obtained from male horses in France, Sweden and Switzerland gave no evidence of a sex-related difference in the genomic DNA. Genomic DNA from 11 streptomycin (STM)-susceptible isolates obtained in Sweden and Switzerland were classified into four genotypes by PFGE. Each of the six genotypes determined among the 17 isolates from these two countries had single phenotypes for resistance or susceptibility to STM.

[Local and systemic infections with bacteria of group EF-4 in dogs, cats and in a badger: bacteriologic and pathologico-anatomic results]. / Lokale und systemische Infektionen mit Bakterien der Gruppe EF-4 bei Hunden, Katzen und bei einem Dachs: Bakteriologische und pathologisch-anatomische Befunde.

Bacteria of Group EF-4 were isolated in 7 dogs and in 5 cats from local purulent lesions in areas of natural openings of the head and from bite wounds, from lungs with focal necrotizing pneumonia in 2 additional cats and from internal organs in a badger with septicemia. The organisms grew mostly in pure culture. In conventional biochemical tests, 12 isolates were shown to belong to biovar EF-4a and 3 isolates to EF-4b. Classification as Group EF-4 was confirmed by assimilation tests performed with the commercially available gallery ATB 32 GN. Results of this study indicate that in veterinary medicine, bacteria of Group EF-4 are not only important from an epidemiological point of view, but that they may be more pathogenic for animals than considered hitherto.

[Causes of prenatal foal loss in Switzerland]. / Ursachen pränataler Fohlenverluste in der Schweiz.

In Switzerland during the foaling season 1988 and 1989 the cause of abortion in 60 foals was investigated. Special attention was paid to infections with equine herpesvirus 1 (EHV 1). Diagnosis were based on post-mortem, histopathological, bacteriological and immunofluorescence investigation. The results confirm data from other countries, that EHV 1 is the most prevalent viral (20%) cause of abortion, followed by various bacterial agents (12%). Other causes were umbilical torsion, twin pregnancy and malformations. In 18% of the cases the investigation of fetuses did not give any results as to the cause of abortion, suggesting maternal factors as only cause of abortion.

[The occurrence of spiral-shaped bacteria in the abomasum of cattle]. / Untersuchungen über das Vorkommen von spiralförmigen Bakterien im Labmagen des Rindes.

The goal of this study was to determine whether Helicobacter pylori or similar bacteria are present in the abomasum of cows. The abomasa of 112 clinically healthy cows were examined at slaughter. Prior to macroscopic examination, samples for bacteriological and histological examination were obtained from the fundus and from the pylorus. Bacteriological examination of the abomasal mucosa included the urease test, the microscopic examination of a Gram's stained smear, and culture on various solid media. Samples from the pylorus (63) were more often positive in the urease test than those from the fundus (35). Examination of Gram's stained smears revealed two groups of suspicious microorganisms; spiral-shaped and rod-shaped bacteria, whereby the latter could not be differentiated morphologically from Helicobacter pylori. Spiral-shaped bacteria were more often isolated from the pylorus (101 samples) than from the fundus (30 samples). The bacteria that resembled Helicobacter pylori were found in seven samples from the pylorus and in seven samples from the fundus. Helicobacter pylori was not cultured in any of the abomasal samples. Tissue samples from the fundus and pylorus were stained with hemalum and eosin and with silver according to Warthin-Starry. All but one abomasum had diffuse infiltration of lymphocytes and plasma cells. Lymphocytic follicles were observed in 109 abomasa. Neutrophils were seen in four abomasa, eosinophils in 37 and parasitic lesions in 20. As in the Gram's stained smears, spiral-shaped and rod-shaped bacteria were seen in silver-stained smears. Spiral-shaped bacteria were found in the pylorus of 96 abomasa and in the fundus and pylorus of one abomasum. The rod-shaped bacteria could not be differentiated from Helicobacter pylori by light microscopy. They were seen in glandular lumina of the superficial region of the mucosa in 97 abomasa. They were limited to the pylorus and fundus in 16 and 59 cases, respectively, and occurred in both these areas in 23 cases. The results of this study indicate that spiral-shaped bacteria may be found frequently in the bovine abomasum. Further investigations are required to determine whether these bacteria are associated with the inflammatory lesions that were observed and whether they play a role in the pathogenesis of abomasal ulcers.

[A case of disseminated intravascular coagulation (DIC) in a cow with endometritis and fetal death]. / Ein Fall von disseminierter intravaskulärer Koagulopathie (DIC) bei einer Kuh mit Endometritis und Fruchttod.

Acute disseminated intravascular coagulation (DIC) was diagnosed in a 3 1/2 year old cow of the Simmental breed. The cow was little less than 6 months pregnant and was admitted to the clinic because of severely disturbed general health. The most important clinical findings were increased heart and breathing rate, rectal temperature of 39.9 degrees C, nosebleed and petechiae on the nasal mucosa. Additionally, the cow showed petechiae on the vaginal mucosa, haemorrhage from the rectum lasting several hours after rectal examination and severe haemoglobinuria. Haematological and biochemical examinations showed increased liver enzymes and severe changes in all coagulation parameters (platelet count, PT, PTT, thrombin time, fibrinogen, fibrin degradation products). Based on the mentioned findings the diagnosis DIC was made. Possible causes were severe necrotic endometritis and placentitis combined with fetal death. High counts of Escherichia coli and Clostridium perfringens were determined in liver, lung and abomasal contents of the aborted fetus as well as in the placenta. Uterine secretion contained Actinomyces pyogenes besides.

[Isolation of Corynebacterium diphtheriae subsp. belfanti from a cow with chronic active dermatitis]. / Isolierung von Corynebacterium diphtheriae subsp. belfanti bei einer kuh mit chronisch-aktiver Dermatitis.

Diphtheria is an acute communicable disease of man caused by C. diphtheriae. Pharyngeal and cutaneous forms are described, where from both toxigenic and nontoxigenic strains can be isolated. The occurrence of C. diphtheriae in dairy cattle has already been reported in the past. The pathogens were isolated from ulcerated teats and from the milk of cows with mastitis as well. These animals were considered to play a role in the transmission of the disease to man. This paper describes the isolation and characterization of C. diphtheriae in a 4 years old cow with generalized, partly ulcerative and purulent skin lesions. Bacteriological examination revealed the presence of very numerous corynebacterium-like organisms, which were characterized as C. diphtheriae subsp. belfanti, a nontoxigenic subspecies of C. diphtheriae. The significance of C. diphtheriae in veterinary medicine and the possible role of cattle as a reservoir of these organisms are discussed.

[Abortion in small ruminants in Switzerland: investigations during two lambing seasons (1996-1998) with special regard to chlamydial abortions]. / Aborte beim kleinen Wiederkäuer in der Schweiz: Untersuchungen während zwei Ablammperioden (1996-1998) unter besonderer Beachtung des Chlamydienabortes.

Abortion cases of 144 goats und 86 sheep were investigated etiologically during 2 lambing seasons (1996/1997, 1997/1998). Macroscopic inspection of fetus and placenta was completed by histopathology and bacteriological isolation of agents. In addition, immunohistologically the following antigens were labeled in formalin-fixed and paraffin-embedded tissue sections: Toxoplasma gondii, Neospora caninum, Chlamydophila abortus (formerly Chlamydia psittaci serovar 1) and Border Disease Virus. From farms with abortions caused by Chlamydophila abortus specific data were recorded. In 75% of abortion cases in sheep and in 59% of cases in goats an etiologic diagnosis could be substantiated. Chlamydophila abortus is the most commonly involved agent in the etiology of caprine and ovine abortion (sheep 39%, goats 23%), followed by Toxoplasma gondii (sheep 19%, goats 15%) and Coxiella burnetti (sheep 1%, goats 10%). All other agents are of minor importance. An infectious cause of abortion based on histopathologic findings without isolation of agents was observed in sheep (10%) and goats (21%). Malformation occurred in sheep (2%) and goats (3%) and lesions suggestive for Vitamin E/Selenium deficiency were seen in goats only (2%).

[Purulent osteomyelitis in fattening pigs]. / Eitrige Osteomyelitis beim Mastschwein.

Purulent osteomyelitis caused by Arcanobacterium pyogenes was diagnosed in three pigs aged between 3 and 4 months by radiological and pathological findings. Osteomyelitis was localized in metaphysis and/or epiphysis of limb bones. The prevalence of osteomyelitis in swine seems to be underestimated because inspection of limb bones is not a routine procedure either at slaughter or at necropsy. Osteomyelitis may also have consequences for meat cutting. Osteomyelitis can be controlled by prophylactic procedures.

[Causes of death and disease in kittens]. / Todes- und Erkrankungsursachen bei Katzenwelpen.

Post-mortem examinations were carried out on 168 kittens 0 to 12 weeks of age. One third of the kittens died of an infectious agent, one third showed non infectious illnesses and in one third the examinations revealed none or vague results.

[Use of a low dose prostaglandin F2 alpha in bitches]. / Anwendung von niedrig dosiertem Prostaglandin F2 alpha bei Hündinnen.

The dosage of Prostaglandin F2 alpha used until the present (100, 250 and 1000 micrograms/kg bw), in order to treat pyometra in the bitch, was accompanied with side effects such as salivation, vomiting and diarrhea. In the present work, the efficiency of low dose Prostaglandin (20 micrograms/kg bw) was examined in two different groups of patients: Group 1: Included 9 bitches pregnant for a period of 5-7 weeks duration. Initially the bitches were treated 3 or 4 times per day with Prostaglandin F2 alpha. In these cases abortion took place within 4 to 11 days. Group 2: 12 dogs, suffering from pyometra, were treated 3 times per day with PGF2 alpha for 8 days. In 9 dogs the pyometra resolved and the bitches came in estrus 2-5 months after treatment. 7 bitches have been mated and 6 of these gave birth to healthy litters. During a follow-up period of at least 10 months there has not been a reoccurrence of pyometra. In 3 out of the 12 dogs the uteri were still enlarged after 8 days of treatment. These bitches underwent ovariohysterectomy and a cystic hyperplasia of the endometrium was diagnosed histologically. The low dose (20 micrograms/kg BW) Prostaglandin F2 alpha induced in all dogs the expulsion of the uterine contents. Side effects during the treatment were not observed.

[Experimental infection of calves and sheep with bovine Giardia isolates]. / Experimentelle Infektion von Kälbern und Schafen mit bovinen Giardia-Isolaten.

9 Giardia-free calves were artificially infected with 1.5-5.1 x 10(6) Giardia cysts originating from Swiss cattle ("bovine isolates"). In 4 of these animals the course of infection was examined. After prepatent periods of 7-8 days all calves excreted high numbers of Giardia cysts for 60-112 days. During patency on 44% of the examination days Giardia cysts and antigen could be detected simultaneously in faecal samples using the flotation method and a sandwich-ELISA, respectively. With the exception of light diarrhoea lasting only for some days at the beginning of patency no other symptoms occurred. Further 5 artificially infected calves were submitted to autopsy. Giardia trophozoites were detected in 4 calves in the jejunum and in 1 animal in the ileum (peroxidase-antiperoxidase method). All animals were simultaneously infected with Campylobacter spp. and/or Rota- and Corona-virus. Electronmicroscopically mucosal attachment sites of Giardia trophozoites had intact microvilli and enterocytes. In various parts of the intestine blunting and flattening of the villi and cellular infiltrations of the mucosa were present. These alterations in calves are generally associated with bacterial and/or viral infections of calves. A Swiss bovine Giardia cyst-isolate was transmitted to 4 Giardia-free conventionally maintained lambs which excreted Giardia cysts after prepatent periods of 10-21 days for 31-61 days.

[Studies of facilities with increased abortion rates in cattle]. / Untersuchungen in Betrieben mit gehäuftem Verwerfen beim Rind.

Cases of abortions in cattle were investigated using several different laboratory procedures. The purpose was to collect information on an individual animal and on a herd basis that would allow a correct etiological diagnosis and also the institution of prophylactic measures. The cause of the abortion was diagnosed in 30% of the cases and in 16.3% of the herds. Due to the complexity of the problem, there is no routine diagnostic procedure that can universally be recommended and applied. There was no association between the number of tests performed or the number of herd mates included in the diagnostic work-up and the diagnostic success rate. In order to make a more efficient use of the diagnostic procedures, a better understanding of the mechanisms leading to abortion, on an individual animal basis and on a herd basis, is required.

[Pestivirus as causative agent of abortion and perinatal mortality in cattle and sheep in Switzerland]. / Ursächliche Beteiligung von Pestiviren an Aborten und perinatalen Todesfällen bei Rindern und Schafen in der Schweiz.

The causal involvement of bovine viral diarrhoea virus (BVDV) and border disease virus (BDV) infection in bovine and ovine abortion and perinatal mortality remain unclear. From 1992 until 1994, 213 bovine and 31 ovine foetuses as well as 36 calves and 25 lambs which had died perinatally were investigated. Tissue samples were tested for the presence of pestiviruses and serum or fluid from the body cavities were analysed for the presence of pestivirus antibodies. Detection of pestiviruses was performed by (i) cell culture isolation, (ii) antigen ELISA and (iii) immunohistochemical staining for viral antigen. For antibody-testing an indirect ELISA was used. In nine bovine foetuses and in two calves BVDV was isolated. Pestiviruses, most likely BDV were detected in one ovine foetus and three lambs. In 6% of the bovine and 11% of the ovine foetuses anti-pestivirus antibodies were detected. However, clinical features and history of bovine cases did not show a correlation with the diagnostic results, In contrast, the presence of central nervous system signs in neonatal lambs and the detection of BDV was correlated.