Enteritis and Septicemia in a Horse Associated With Infection by Escherichia fergusonii

This report presents a clinical case of enteritis and septicemia with subsequent death of an adult horse associated with Escherichia fergusonii, in Germany. The patient died within 24 hours after onset of clinical signs, including fever and erosive stomatitis. Major pathologic findings were severe, acute, fibrinous, and necrotizing enteritis; severe, acute, fibrinous endocarditis; as well as multiple, hyaline thrombi in blood vessels. E fergusonii and Escherichia coli were isolated in moderate to high amounts from the intestine, the tricuspid valve, and the liver. This article represents the first case report of lethal enteritis and septicemia associated with E fergusonii infection in a horse.     

Multiplex PCR assay for detection of Clostridium perfringens in feces and intestinal contents of pigs and in swine feed

A multiplex polymerase chain reaction (PCR) assay, developed to detect the alpha-toxin and enterotoxin genes (cpa and cpe, respectively) of Clostridium perfringens, was used to identify enterotoxigenic isolates of this organism from feces and intestinal contents of pigs and from feed samples from pig farms in Iowa. The organism was grown on tryptose-sulfite-cycloserine (TSC) agar, TSC agar without egg-yolk, sheep blood agar, or in brain heart infusion broth or cooked meat medium. DNA was extracted by boiling and the PCR assay was carried out using reagents from a commercial kit. The 319 bp amplification product of cpa and the 364 bp product of cpe were visualized under UV light after electrophoresis in a 2% agarose gel containing ethidium bromide. The average sensitivity of the assay, determined on artificially contaminated feces, was 9.2×10⁴ colony forming units per gram. Assay of 97 isolates from feces and intestinal contents revealed cpa in 89, but all were negative for cpe. While 28% of the 442 total samples cultured yielded C. perfringens, only 5% of 298 fecal or intestinal contents samples were positive upon direct examination by the PCR assay. Ninety-one and eight-tenths % of isolates with the phenotype of C. perfringens were cpa positive by PCR. Forty-three percent of feed samples were culture positive, while 48.3% were PCR positive for cpa. None of these were cpe positive. We conclude that PCR is a useful assay for rapid detection of C. perfringens in feed, and for confirmation of the identity of isolates presumed to be C. perfringens.     

Application of a new diagnostic approach to a bovine genital campylobacteriosis outbreak in a Saskatchewan beef herd

A new real-time quantitative polymerase chain reaction (qPCR) test was used to diagnose Campylobacter fetus subsp. venerealis infection associated with dramatic reproductive losses in a commercial cow-calf herd. The results were verified with repeated culture, phenotypic characterization of the organism and DNA sequencing. This case demonstrates the need for a practical field test for C. fetus subsp. venerealis and the importance of considering this organism as a potential cause of pregnancy failure in beef herds.     

Natural history of Ehrlichia ruminantium

Ehrlichia ruminantium is an obligately intracellular proteobacterium which causes a disease known as heartwater or cowdriosis in some wild, and all domestic, ruminants. The organism is transmitted by ticks of the genus Amblyomma, and it is of serious economic importance wherever the natural vectors occur, an area which includes all of sub-Saharan Africa, and several islands in the Caribbean. The disease was first recognized in South Africa in the 19th century, where its tick-borne nature was determined in 1900, but the organism itself was not demonstrated until 1925, when it was recognized to be a rickettsia, initially named Rickettsia ruminantium. It was thus the first species of what are now known as Ehrlichia to be discovered, and most of the early work to elucidate the nature of the organisms, and its reservoirs and vectors, was performed in South Africa. The next milestone was the development, in 1945, of an infection and treatment regimen to immunize livestock, and this is still the only commercially available “vaccine” against the disease. Then in 1985, after fruitless attempts over many years, the organism was propagated reliably in tissue culture, opening the way for the first application of the newly developed techniques of molecular genetics. From 1990 onwards the pace of heartwater research accelerated rapidly, with notable advances in phylogeny, diagnosis, epidemiology, immunology, and vaccine development. The complete genome sequence was published in 2005, and during the last two years a new understanding has arisen of the remarkable genetic variability of the organism and new experimental vaccines have been developed. Despite all this the goal of producing an effective vaccine against the disease in the field still remains frustratingly just beyond reach. This article summarises our current understanding of the nature of E. ruminantium, at a time when the prospects for the development of an effective vaccine against the organism seem better than at any time since its discovery 83 years ago.     

Salmonella Antimicrobial Activity of Selected Strains of Enterolactobacillus Species Isolated from the Gastrointestinal Tract of the Horse

The gastric mucosa and the mucosa of the right and left dorsal colon were biopsied in each of the 15 horses, and a total of 45 samples were collected. Mucosal samples were cultured using a Lactobacillus enrichment broth. While numerous Lactobacillus strains were identified, Lactobacillus reuteri was the most common organism identified. Sixteen strains of Lactobacillus reuteri were selected for antimicrobial testing. Salmonella antimicrobial activity was identified in six out of 16 strains tested. Organisms with Salmonella antimicrobial activity were cultured from the stratified squamous epithelium of the stomach and the mucosa of the right and left dorsal colon.     

Diagnosis of proliferative enteritis in frozen and formalin-fixed,paraffin-embedded tissues from a hamster,horse, deer and ostrich using a Lawsonia intracellularis-specific multiplex PCR assay

Proliferative enteritis (PE) is an enteric disease that has been reported in a variety of animals. It is caused by an obligate intracellular bacterium identified in swine as Lawsonia intracellularis. The organism can be detected ante-mortem in swine with PE using molecular diagnostic methods. The disease can be diagnosed post-mortem in all species by gross examination of tissues and special histologic staining procedures. In this study we extracted total DNA from frozen or formalin-fixed, paraffin-embedded tissues from cases of pig, hamster, horse, deer and ostrich PE. The samples were subjected to a multiplex PCR reaction using primers specific for a swine isolate of L. intracellularis. Identical sized PCR products were detected in samples from all animals with PE and the specificity of the PCR reaction for L. intracellularis was demonstrated by Southern-blotting and hybridization using specific probes. These results suggest that the intracellular organism of PE in these species are all very closely related to the causative agent of PE in swine, L. intracellularis. In addition, this multiplex PCR assay can be used to detect the organism in frozen or archival tissues, facilitating retrospective diagnosis of PE.     

Development of a loop-mediated isothermal amplification assay for the detection of Mycobacterium bovis

Bovine tuberculosis caused by Mycobacterium bovis is an important zoonosis. In this study, a simple, rapid method for detecting this organism was developed based on loop-mediated isothermal amplification of the mpt83 gene. The technique will be of value in the clinical and field-based diagnosis of M. bovis and can differentiate it from other bacteria such as Corynebacterium diphtheriae, Streptococcus pneumoniae, β-haemolytic streptococci, Pseudomonas aeruginosa, Yersinia pseudotuberculosis, Staphylococcus aureus, and Klebsiella pneumoniae. The limit of detection was 10 copies/μL and the results were corroborated by PCR. The method was highly specific and more sensitive than PCR.     

Bacteroides ruminicola as a possible cause of “lumpy-jaw” in Bennett's wallabies

Bacteroides ruminicola subspecies brevis was isolated from the jaw lesions of three Bennett's wallabies with “lumpy-jaw”. Nocardia was not isolated from any of the three animals. It is suggested that Bacteroides ruminicola subspecies brevis is the causal organism of “lumpy-jaw” and was one of the organisms noted in this condition by Fox in 1923. As far as we are aware this subspecies has not previously been reported as a pathogen in animals or human beings. The results of antibiotic sensitivity tests are given and therapy with metronidazole or clindamycin is suggested.     

PCR-based detection of genes encoding virulence determinants in Staphylococcus aureus from bovine subclinical mastitis cases

The present study was carried out to genotypically characterize Staphylococcus aureus (S. aureus) isolated from bovine mastitis cases. A total of 37 strains of S. aureus were isolated during processing of 552 milk samples from 140 cows. The S. aureus strains were characterized phenotypically, and were further characterized genotypically by polymerase chain reaction using oligonucleotide primers that amplified genes encoding coagulase (coa), clumping factor (clfA), thermonuclease (nuc), enterotoxin A (entA), and the gene segments encoding the immunoglobulin G binding region and the X region of protein A gene spa. All of the isolates yielded an amplicon with a size of approximately 1,042 bp of the clfA gene. The amplification of the polymorphic spa gene segment encoding the immunoglobulin G binding region was observed in 34 isolates and X-region binding was detected in 26 isolates. Amplification of the coa gene yielded three different products in 20, 10, and 7 isolates. The amplification of the thermonuclease gene, nuc, was observed in 36 out of 37 isolates. All of the samples were negative for the entA gene. The phenotypic and genotypic findings of the present strategies might provide an understanding of the distribution of the prevalent S. aureus clones among bovine mastitis isolates, and might aid in the development of steps to control S. aureus infections in dairy herds.     

Some problems associated with the identification of Corynebacterium equi

One hundred strains of Corynebacterium equi were examined using standard laboratory procedures. Variations in colonial morphology were noted. The ability to reduce nitrate and to produce urease was shown to be valuable for confirmation of identification of C. equi. Production of hydrogen sulphide was a variable characteristic of C. equi. It is concluded that for those situations where the source of the organism gives no clue as to its identify, positive identification of C. equi may be difficult because of variability in colonial appearance and biochemical reactions.     

Brucella abortus infection in indigenous Korean dogs

Three dogs reared on a dairy farm with a high incidence for Brucella abortus were serologically positive for B. abortus and no other Brucella spp. The identity of the organism was confirmed to be B. abortus by AMOS (abortus melitensis ovis suis)-polymerase chain reaction with specific primers for B. canis. One hundred percent homology of the canine isolate and the bovine pathogen isolated from the farm was demonstrated. The only possible source of infection was infected cattle on the same farm. It is suggested that dogs be routinely included in brucellosis surveillance and eradication programs.     

Studies ofStreptococcus suis type 2 infection in pigs

Investigations into the immunology, pathogenesis and epidemiology ofStreptococcus suis type 2 infections were carried out in experimental pigs and in naturally-occurring field outbreaks of disease.The capsular polysaccharide fromStr. suis type 2 was shown to induce opsonic antibodies in pigs when injected with Freund's incomplete adjuvant, but difficulties encountered in experimental production of the disease prevented a study of their protective effects.Problems with the bactericidal tests led to an investigation of other assays for antibodies againstStr. suis type 2, namely, a phagocytic test with pig neutrophils, a mixed reverse passive antiglobulin haemagglutination test and an indirect haemagglutination test. There was evidence that with modifications both the latter tests would be useful.Transmission studies in 39 conventionally-reared and 7 hysterectomy-derived, colostrum-deprived pigs yielded interesting results with regard to the distribution of the organism in relation to the disease process.Tonsil carriage in clinically-healthy pigs was demonstrated after experimental and natural infection. Detectable carrier rates varied between 0 and 59%. The organism was shown to persist in the presence of circulating opsonic antibodies and in pigs on penicillin-medicated feed. Attempts to isolate the organism from the genital tract were unsuccessful.Medicated early weaning and classical SPF techniques applied to infected herds appeared to be effective in producing pigs free fromStr. suis type 2 infection.     

Establishment of a Salmonella-Free Guinea Pig Colony

Salmonellosis due to Salmonella typhimurium was enzootic in a guinea pig breeding colony for over 25 years. A Salmonella-free auxiliary colony was established by removing weanlings from the infected colony to a clean area, and preventing infection. Examination of agglutinin titers and necropsy specimens indicated that the auxiliary colony was still free from Salmonella 18 months after its establishment while 24% of the guinea pigs dying in the infected colony yielded Salmonella typhimurium.     

Natural History of Ehrlichia chaffeensis: Vertebrate hosts and tick vectors from the United States and evidence for endemic transmission in other countries

Ehrlichia chaffeensis, an intracellular gram-negative zoonotic bacterium, is the causative agent of human monocytotropic ehrlichiosis (HME). In humans, the disease can range from a mild, non-specific illness with few to no clinical signs to a moderately severe to fatal disease, especially those with compromised immune systems. E. chaffeensis is maintained in a complex cycle involving white-tailed deer (WTD; Odocoileus virginianus) as a primary reservoir and the lone star tick (LST; Amblyomma americanum) as a primary vector. Numerous other species are naturally exposed to E. chaffeensis and disease has been documented in some domestic animals and wildlife including domestic dogs and ring-tailed lemurs. The organism has been found throughout the natural range of the LST and as the tick continues to expand its range, the geographic range of risk for E. chaffeensis infections will likely continue to expand. Recent data have indicated that E. chaffeensis, or a closely related organism, has been found in many species of ticks and vertebrate hosts in numerous countries.     

Haemophilus somnus-induced Otitis and Meningitis in a Heifer

The purpose of this report is to describe the lesions of otitis and meningitis associated with Haemophilus somnus infection in a heifer.

The animal was found dead in its pen. At necropsy, there was a severe fibrinopurulent meningitis and suppurative otitis media and interna. The external ear canal contained inspissated debris. Haemophilus somnus was recovered in pure culture from the meningeal exudate. The identity of the organism was confirmed by fluorescent antibody and agar gel immunodiffusion tests. Possible pathogenetic mechanisms are discussed.

     

Helicobacter spp. in the saliva,stomach, duodenum and faeces of colony dogs

The role of Helicobacter spp. infection in canine gastrointestinal disease is unclear and routes of transmission are of epidemiological and zoonotic importance. The aim of this study was to identify Helicobacter spp. in the saliva, stomach, duodenum and faeces of dogs using a multiplex PCR, and to evaluate any attendant histopathological changes. Helicobacter canis was the most common species detected in saliva and faeces and no correlation between the presence of Helicobacter spp. and histopathological changes in either the stomach or duodenum was observed. All dogs examined were co-infected with up to four species of the organism. This is the first time these bacteria have been studied at species level at multiple sites within the canine alimentary tract.     

Prevention of Salmonella contamination of finished soybean meal used for animal feed by a Norwegian production plant despite frequent Salmonella contamination of raw soy beans, 1994–2012

Background

Salmonella contaminated animal feed is a major source for introducing Salmonella into the animal derived food chain. Because soybeans frequently are contaminated with Salmonella, soybean meal used as animal feed material, a by-product of a “crushing plant” which produces oil from soybeans, can be important source of Salmonella in the animal feed.We report the successful control of Salmonella from 1994 to 2012 in a Norwegian crushing plant producing soybean meal from imported soy beans. The results are based on an officially supervised HACCP based program including annual testing of around 4000 samples.

Results

During the 19-year period, 34% of samples collected during unloading of ships delivering soybeans yielded Salmonella; the proportion of samples from ships that yielded Salmonella varied from 12-62% each year. Dust samples from all shiploads from South America yielded Salmonella. In total 94 serovars of Salmonella were isolated, including nine (90%) of the EU 2012 top ten serovars isolated from clinical cases of salmonellosis in humans, including major animal pathogenic serovars like Spp. Typhimurium and Enteritidis.The effectiveness of the HACCP based control was indicated by a low prevalence of Salmonella contamination in the clean area of the plant, which is considered to be the main reason for the successful prevention of Salmonella in the end product. Despite extensive testing, no sample from the finished soybean meal product was found to be Salmonella contaminated.

Conclusions

This study shows that a HAACP-based control program in a soybean crushing plant can produce Salmonella free soybean meal despite frequent Salmonella contamination of raw soybeans. That approach is suggested as an effective way to minimize the risk of Salmonella exposure of the animal feed mills and contamination of the subsequent animal feed chain.     

Utility of contrast-enhanced ultrasound in differential diagnosis of adrenal tumors in dogs

This prospective case study aimed to clarify the clinical significance of contrast-enhanced ultrasound (CEUS) for the differential diagnosis of canine adrenal tumors. Forty-three client-owned dogs with adrenal tumors were included. All dogs underwent CEUS, which was evaluated qualitatively and quantitatively. The peak signal intensity (PI), time to peak signal intensity (TPI), mean transit time (MTT), upslope, and downslope were calculated for each time-intensity curve. The histopathological diagnosis of each resected mass was compared with the CEUS findings and parameters. Enhancement distribution, vascularity, tortuous nourishing vessels, enhancement pattern, and late-phase enhancement did not differ significantly between adrenal cortical adenoma (CA), adenocarcinoma (CAC), and pheochromocytoma (PHEO) in qualitative assessment. In PHEO, the TPI was significantly more rapid compared with that in CA (P=0.0287) and CAC (P=0.0404). The MTT in PHEO was significantly shorter than that in CA (P=0.0016) and CAC (P=0.0003). Upslope in PHEO was larger than that in CAC (P=0.0406). Downslope in PHEO was significantly larger than that in CA (P=0.0048) and CAC (P=0.0018). A receiver operating characteristic curve analysis demonstrated that the area under the MTT curve yielded 0.91 for distinguishing PHEO from adrenocortical tumors in dogs; an MTT cut-off value less than 6,225 msec yielded a sensitivity of 69%, specificity of 94%, and likelihood ratio of 12.46. CEUS appears to be clinically applicable for the differential diagnosis between cortical and medullary origins of primary adrenal tumors in dogs.     

Serological investigation of Corynebacterium pseudotuberculosis infection in sheep--correlation between the hemolysis inhibition test and the ELISA test

Corynebacterium pseudotuberculosis causes caseous lymphadenitis in sheep and goats. The animals may be infected without showing clinical symptoms. Several serotests have therefore been employed to detect infected animals. Shen et al (1982) performed an enzyme linked immunosorbent assay (ELISA) to detect antibodies against the organism using cell wall antigens. Maki et al (1985) found that the toxin of the bacterium was a better antigen for assessing infection in the ELISA test. They reported that the antitoxin ELISA appeared to be as sensitive as the antihemolysin inhibition test (Zaki 1968).