Fatal suppurative meningoencephalitis caused by Klebsiella pneumoniae in two calves

One calf died (No. 1) and another was euthanized following astasia (No. 2). Histopathological examination revealed suppurative meningoencephalitis in these calves. Klebsiella pneumoniae antigens were detected in lesions. Thymocytes were decreased in the thymus cortex in both cases. 16S rRNA gene sequencing of the No. 1 isolate and bacterial extracts from formalin fixed paraffin embedded sections of No. 2 revealed that both samples were K. pneumoniae. The No. 1 isolate showed multidrug resistance against penicillin antibiotics, fosfomycin, streptomycin, macrolide antibiotics, tetracycline antibiotics, and clindamycin. Immunosuppression is a significant septicemic K. pneumoniae infection risk factor. Our study provides new aspects regarding K. pneumoniae infections in cattle, bacterial meningoencephalitis differentiation, and K. pneumoniae and bacterial meningoencephalitis treatments.     

Septicemic salmonellosis in a two-humped camel calf (<Emphasis Type="Italic">Camelus bactrianus</Emphasis>)

A 1-week old, two-humped female camel (Camelus bactrianus) calf with continual whining, epiphora, anorexia, muscle twitching, and lateral recumbency was referred to a veterinary hospital. Although she died shortly after preliminary clinical examination, but necropsy was performed and tissue samples were taken for further microbiological and pathological examinations. On bacteriological investigation, Salmonella typhimurium and Streptococcus agalactiae were isolated. Histopathologically, lesions consisted of hyperemia and hemorrhage in all serosal and mucosal surfaces, gastroenteritis, and purulent ascites, associated with suppurative omphalitis. Acute nutmeg liver demonstrated centrilobular congestion and moderate fatty changes without any inflammatory cell infiltration. The abomasal and intestinal mucosa were hemorrhagic and erosive. The brain was hyperemic with severe fibrinopurulent meningoencephalitis. Except for dromedary camels and llamas, there has been no previous report of an acute, fatal septicemia in a two-humped camel calf due to S. typhimurium accompanied by S. agalactiae.     

Phenotypic and genotypic characterization of Actinobacillus suis sensu stricto isolated from a dairy calf

The species of the genus Actinobacillus have so far been associated with specific animal hosts, and A. suis sensu stricto, an opportunistic pathogen of swine, is rarely isolated from ruminants. We describe here the isolation of A. suis sensu stricto from a newborn calf that died on a dairy farm in Japan. Identification of the isolate was performed by phenotypic and genotypic characterization, with the latter consisting of nucleotide sequence analyses of the 16S rRNA gene plus three housekeeping genes, rpoB, infB and recN.     

Isolation and characterization of a bovine isolate of Neospora caninum with low virulence

Neospora caninum tachyzoites were isolated from the brain of an asymptomatic naturally infected calf with precolostral-specific antibodies. The new isolate, named Nc-Spain 1H, was identified as a member of the N. caninum species based on its internal transcribed spacer 1 (ITS-1) sequence and was genetically characterized using microsatellite markers. Multilocus analysis showed that Nc-Spain 1H was genetically different from other N. caninum isolates. We compared the in vitro tachyzoite yield and viability rate of the Nc-Spain 1H and Nc-1 isolates in a plaque assay. The lower tachyzoite yields displayed by Nc-Spain 1H were complemented with a significantly lower viability rate. Moreover, in an in vitro tachyzoite–bradyzoite stage conversion assay, the percentage of Nc-Spain 1H bradyzoite conversion was similar to that of the cystogenic isolate Nc-Liv, with the exception that Nc-Spain 1H produced only intermediate bradyzoites. The pathogenicity of Nc-Spain 1H was examined in BALB/c mice, and the results demonstrated that Nc-Spain 1H failed to induce clinical signs or mortality and that no parasite DNA was detected in the brain during the chronic stage of infection. In a pregnant mouse model, Nc-1 infection resulted in high transplacental transmission, leading to a high neonatal mortality rate over time. In contrast, the offspring survival rate from Nc-Spain 1H-infected dams was almost 100%, and N. caninum DNA was detected in only one pup. These data show that Nc-Spain 1H appears to be a low virulence isolate and may be a suitable candidate for live vaccine development.     

Campylobacter jejuni abortions in two beef cattle herds in Saskatchewan

Abortions, accompanied by placental retention and weight loss, occurred during February and March in 19% of 120 and 10% of 108 beef cows and heifers on two neighboring ranches in southern Saskatchewan. A diagnosis of Campylobacter jejuni abortion was made based on lesions of necrotizing and suppurative placentitis and fetal bronchopneumonia in association with the culture of large numbers of C. jejuni from placentas and fetal tissues.

Campylobacter jejuni was isolated with variable frequency from fecal samples of aborting and healthy cows, and scouring and healthy calves. Campylobacter jejuni serotype 2 (Lior) was isolated from fetal tissues and feces of a scouring calf, whereas C. jejuni serotypes 1, 4, 5 and 99 were isolated from feces of in-contact cattle. We hypothesized that the source and mode of transmission of C. jejuni was fecal contamination of water supplies and feeding grounds by carrier cows or wildlife.

     

Genetic and antigenic analysis of Chlamydia pecorum strains isolated from calves with diarrhea

Chlamydia pecorum (designated 22–58) was isolated in 2010 in HmLu-1 cells from the jejunum of a calf which died of necrotizing enterocolitis in Yamaguchi Prefecture, Japan. Immunohistochemical staining identified C. pecorum positive reactions in the jejunal villi. C. pecorum, designated 24–100, was isolated from the feces of a calf with diarrhea in another farm in Yamaguchi Prefecture in 2012. A significant increase in neutralizing antibody titers against C. pecorum was confirmed in paired sera. Nucleotide sequence identities of omp1 genes of the 2 isolates were 100%. The isolates were genetically and antigenically more closely related to C. pecorum Bo/Yokohama strain isolated from cattle with enteritis in Japan than to the other prototype strains, Bo/Maeda isolated from cattle with pneumonia and Ov/IPA isolated from sheep with polyarthritis. These results indicate that C. pecorum strains similar to 22–58 and 24–100 might be endemic in Yamaguchi Prefecture and cause enteric disease in cattle.     

Isolation of pathogenic strains of Haemophilus somnus from the female bovine reproductive tract.

The prevalence of Haemophilus somnus in the genital tract of slaughtered and live cows in southern Ontario was investigated. The vagina and uterus of slaughtered cows were swabbed separately. Live cows were examined and sampled in two field surveys: Centre A and Centre B. In the former, aspirated mucus secretions and in the latter, specimens obtained by guarded swabbing were examined bacteriologically. Haemophilus somnus was isolated from 28 genital tracts of 461 slaughtered (6.1%), and seven of 199 live (3.5%) cows during the centre B survey. The isolates were recovered from both normal and diseased reproductive tracts. Fourteen strains isolated from genital organs were examined for pathogenicity in vivo to test the occurrence of pathogenic isolates. In the initial stage of the in vivo study on pathogenicity, each of the fourteen isolates was examined on one calf using an intracisternal inoculation. Subsequently, one pathogenic and one nonpathogenic strain were inoculated into five calves each to statistically confirm their pathogenic potential. Of 14 genital isolates of H. somnus examined in an intracisternal calf assay, six (43%) caused a fatal peracute neurological disease, while eight were nonpathogenic. A comparative pathological study of pathogenic and nonpathogenic isolates showed that the former caused a severe fatal suppurative meningoencephalitis whereas the latter caused no lesions whatsoever or a mild leukocytic leptomeningitis. The salient data obtained in this study indicate that there are pathogenic strains of H. somnus in the genital tract of apparently normal cows as well as of those with inflammatory disease.     

Screening for Indian Isolates of Predacious Fungi for Use in Biological Control against Nematode Parasites of Ruminants

Four isolates of predacious fungi, two each of Arthrobotrys oligospora isolated from a sheep and a male crossbred calf and of Duddingtonia flagrans isolated from a sheep and a female buffalo in western India, were studied for their suitability as biocontrol agents against parasitic nematodes of ruminants, using growth assay, predatory activity, germination potential and ability to survive passing through the ruminants gut as criteria. The study showed that isolates of D. flagrans grew well in artificial media, had encouraging predatory activity, produced profuse chlamydospores that germinated easily at 25°C and could survive passage through the ruminant gut. The ovine isolate of D. flagrans was superior in all respects to the isolate from buffalo and was the most promising candidate for biological control of nematode parasites of ruminants.     

Phylogenetic characterization of Isospora jaracimrmani oocysts from a veiled chameleon (family Chamaeleonidae; Chamaeleo calyptratus) reared at a zoo in Ishikawa,Japan

Oocysts of Isospora sp. were detected in the feces of a veiled chameleon (family Chamaeleonidae; Chamaeleo calyptratus) kept at a zoo in Ishikawa, Japan. Phylogenetic analysis placed the sequence in the cluster of Isospora spp. isolated from reptiles. Based on a comparison of morphological data of ten previously reported Isospora species from the Chamaeleonidae family, this isolate was morphologically similar to I. jaracimrmani, which has been considered to be a virulent species. This case study suggests the possibility that species of Isospora might not always cause disease because the animal that shed these oocysts showed no symptoms for more than two months.     

Rickettsia lusitaniae sp. nov. isolated from the soft tick Ornithodoros erraticus (Acarina: Argasidae)

In this study a novel Rickettsia from the spotted fever group, isolated from Ornithodoros erraticus soft ticks collected from pigpens in the south of Portugal, is described. After initial screening revealed Rickettsia-positive ticks, isolation attempts were then performed. Successful isolates were achieved by shell-vial technique using Vero E6 cells at 28 °C. Molecular characterization of the isolate was performed based on analysis of five rickettsial genes gltA, ompA, ompB, sca1 and htr with their subsequent concatenation along with other rickettsial species resulting in a clustering of the new isolate with Rickettsia felis and Rickettsia hoogstraalii. The degree of nucleotide sequence similarity with other rickettsiae fulfills the criteria for classification of our isolate as a novel species. The name Rickettsia lusitaniae sp. nov. (= CEVDI PoTiRo) is proposed for this new species found in O. erraticus.     

Single nucleotide polymorphisms in the bovine Histophilus somni genome; a comparison of new and old isolates

Histophilus somni, a causative agent of the bovine respiratory disease complex, can also cause a variety of systemic disorders, including bronchopneumonia, myocarditis, pericarditis, arthritis, pleuritis, and infectious thrombotic meningoencephalitis. The purpose of this study was to determine if currently circulating strains differ from those of the 1980s by identifying genomic changes. Single nucleotide polymorphisms (SNPs) and insertion and deletion (INDEL) sites were examined by whole-genome sequencing in 12 samples, 6 old and 6 new. The 31 028 SNP/INDELs recorded were compared against the reference genome sequence of the pathogenic H. somni strain 2336. The distribution of about 75% of these SNPs within a specified gene differed between old and new isolates and did not follow any particular pattern. The other 25% clustered into 2 groups containing the same SNPs in various genes: group I included 5 old isolates and 1 new isolate; group II included 5 new isolates and 1 old isolate. For putative virulence genes there were more SNPs in group I compared with strain 2336, itself an older isolate, than in group II. Although only 25% of all the SNPs formed 2 clusters, the results suggest some genetic difference in various genes between old and new strains.     

Analysis of Salmonella enterica serotype Enteritidis isolated from human and chickens by repetitive sequence-PCR fingerprinting, antibiotic resistance and plasmid profiles

A total of 22 Salmonella enterica serotype Enteritidis (S. Enteritidis) strains isolated from human and chicken were subjected to DNA fingerprinting by repetitive sequence PCR using ERIC and BOX primers, antibiotic resistance and plasmid patterns. Both ERIC and BOX PCR amplification data revealed a highly genetic homogeneity between isolates from human and chicken except one isolate, which originated from chicken and showed a different DNA band pattern from others. Eleven of 22 S. Enteritidis isolates (50%) were resistant to more than one antibiotics and characterized by 5 resistance patterns. The most common pattern was penicillin resistant (63.6%). Only one isolate from chicken showed a multiple drug resistance patterns to 4 antibiotics. All 22 S. Enteritidis isolates harbored more than two plasmids with eight different plasmid profiles including two to six plasmids with approximate molecular size ranging from 1.9 to 21 kb. A band of 15 kb size was detected in all isolates tested, however, the band sizes smaller than 15 kb were found only in isolates from chicken.     

Genotyping of Streptococcus agalactiae strains isolated from fish,human and cattle and their virulence potential in Nile tilapia

Streptococcus agalactiae (Lancefield group B; GBS) is a pathogen that causes meningoencephalitis in fish, mastitis in cows, and neonatal sepsis in humans. The objective of this study was to characterize S. agalactiae isolated from fish (n = 27), cows (n = 9), and humans (n = 10) using pulsed-field gel electrophoresis (PFGE) and to investigate the virulence of the identified strains in Nile tilapia (Oreochromis niloticus). The PFGE types were determined by dendogram analyses and the in vivo virulence was evaluated by experimental infection (using i.p. and immersion routes) of Nile tilapia. Among the fish strains, 5 different PFGE patterns were observed and 21 strains showed the same genetic pattern. In some farms two or three profiles occurred simultaneously. The bovine and human strains exhibited high genetic diversity and few relationships were established among S. agalactiae strains from the three host origins analyzed. Eight S. agalactiae strains from fish caused high mortality of Nile tilapia. Three bovine strains infected Nile tilapia (by i.p. route) and two of those strains caused clinical signs of meningoencephalitis. All human strains (n = 5) infected Nile tilapia (by i.p. route) and meningoencephalitis was induced by one strain (by both i.p. and immersion routes). In conclusion, the analyzed strains from the three natural hosts did not show genetic relatedness, yet some of the bovine and human strains were able to infect fish and cause meningoencephalitis. We suggest that genetic linkage is not a prerequisite for S. agalactiae to cross the host-specific barrier.     

Suppurative granulomatous sinorhinitis associated with Nocardia spp. infection in a cat

A 9-year-old spayed female cat was examined for cheek skin drainage. The skin lesion did not respond to medical therapy; thereafter, facial deformity developed. A computed tomography revealed an intranasal mass and maxillary osteolysis. The mass was histopathologically diagnosed as suppurative granulomatous inflammation caused by filamentous bacteria. The lesion responded well to radiation therapy. Although actinomycosis was suspected histopathologically, no actinomycetes were detected in the nasal lesion by a bacterial culture conducted at a commercial laboratory. The submandibular lymph node and subcutaneous tissue exhibited swelling. Microbiological examination and genetic analysis based on 16S rDNA gene sequence revealed that Nocardia spp. were isolated from both lesions.     

Antimicrobial resistance and virulence factors in Escherichia coli from swedish dairy calves

Background

In Sweden, knowledge about the role of enteropathogenic Escherichia coli in neonatal calf diarrhea and the occurrence of antimicrobial resistance in E. coli from young calves is largely unknown. This has therapeutic concern and such knowledge is also required for prudent use of antimicrobials.

Methods

In a case control study Esherichia coli isolated from faecal samples from dairy calves were phenotyped by biochemical fingerprinting and analyzed for virulence genes by PCR. Antimicrobial susceptibility was tested by determination of minimum inhibitory concentration (MIC). Farm management data were collected and Fisher''s exact test and univariable and multivariable logistic regression analysis were performed.

Results

Of 95 E. coli tested for antimicrobial susceptibility 61% were resistant to one or more substances and 28% were multi-resistant. The virulence gene F5 (K99) was not found in any isolate. In total, 21 out of 40 of the investigated virulence genes were not detected or rarely detected. The virulence genes espP, irp, and fyuA were more common in resistant E. coli than in fully susceptible isolates (P < 0.05). The virulence gene terZ was associated with calf diarrhea (P ≤ 0.01).The participating 85 herds had a median herd size of 80 lactating cows. Herds with calf diarrhea problems were larger (> 55 cows; P < 0.001), had higher calf mortality (P ≤ 0.01) and calf group feeders were more in use (P < 0.05), compared to herds without calf diarrhea problems.There was no association between calf diarrhea and diversity of enteric E. coli.

Conclusions

Antimicrobial resistance was common in E. coli from pre-weaned dairy calves, occurring particularly in calves from herds experiencing calf diarrhea problems. The results indicate that more factors than use of antimicrobials influence the epidemiology of resistant E. coli.Enteropathogenic E. coli seems to be an uncommon cause of neonatal calf diarrhea in Swedish dairy herds. In practice, calf diarrhea should be regarded holistically in a context of infectious agents, calf immunity, management practices etc. We therefore advice against routine antimicrobial treatment and recommend that bacteriological cultures, followed by testing for antimicrobial susceptibility and for virulence factors, are used to guide decisions on such treatment.     

牛病毒性腹泻病毒JN株的分离鉴定及E2基因的序列分析

本研究从疑似牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)感染牛的分泌物与排泄物中分离鉴定1株牛病毒性腹泻病毒,并进行E2基因序列分析。结果表明,分离株病毒命名为JN株;Reed-Muench法测定分离株病毒TCID₅₀为10^-7.5/0.1 mL;病毒中和试验结果表明,BVDV JN分离株可被BVDV阳性血清特异性中和,而不能被BVDV阴性血清中和;分离株病毒E2基因序列测序结果表明,该分离毒株属于BVDVⅠa亚型。     

First Report of Methicillin-Resistant Staphylococcus aureus ST5 and ST398 from Purebred Lusitano Horses

Methicillin-resistant Staphylococcus aureus (MRSA) was first described in horses in 1996. The frequency of MRSA colonization in horses varies among European countries, but it is unknown in Portugal. The aim of this study was to screen for MRSA nasal carriage in a sample of horses entering the Equine Unit, Large Animal Veterinary Teaching Hospital of the Faculty of Veterinary Medicine, Lisbon, Portugal. Seventy-one horses were swabbed, and MRSA was identified by selective isolation on a chromogenic medium. Two S aureus isolates showed resistance to oxacillin (minimum inhibitory concentration >4 μg/mL) and contained the mecA gene. Both strains were isolated from purebred Lusitano horses that lived in farms with more than 20 equines. These MRSA strains represented two different clones: isolate FMVA3/10 was an MRSA sequence type ST5 with a staphylococcal cassette chromosome mec VI, coresistant to erythromycin and clindamycin; and isolate FMVA16/10 was sequence type ST398, with a staphylococcal cassette chromosome mec IV, coresistant to tetracycline, gentamicin, and trimethoprim. Isolate FMVA3/10 represents a human epidemic clone not previously reported among horses in Europe, which once again reinforces the fact that transmission of MRSA clones between horses and humans occurs. Isolate FMVA16/10 represents the first report of the detection of MRSA ST398 among horses in Portugal. Lusitano horses can carry animal and human MRSA in the nostrils, acting as reservoirs, which can potentially be transmitted to humans.     

Salmonella dublin Septicemia in Two Puppies

Two eight week old purebred female Bull Terrier puppies died within 24 hours of each other as a result of a septicemia caused by Salmonella dublin. The salient clinical features were: temperature of 41°C; rapid breathing; fluid, blood-stained stools; prostration and death. Pathological findings included embolic pneumonia, splenitis, myocarditis, nephritis and meningoencephalitis. Salmonella dublin was isolated from the spleen, lung and kidneys of both puppies.     

Disruption of blood-brain barrier by an Escherichia coli isolated from canine septicemia and meningoencephalitis

Escherichia coli (E. coli) is one of the common pathogenic bacteria in veterinary clinical infection. As an opportunistic microorganism, E. coli normally does not cause diseases. However, it causes infections under certain circumstance to domesticated animal and poultry, resulting in severe diarrhea, septicemia, and respiratory infections. Although there are increasing reports regarding the infections of E. coli to domestic animals and poultry, the infection of E. coli in dogs is relatively less reported, especially on septicemia and meningoencephalitis. Here, we reported the isolation and identification of an E. coli isolate named CEC-GZL17 from dogs characterized by septicemia and sudden death, and found that CEC-GZL17 is able to cause meningoencephalitis. Exploration on the potential mechanism underlying meningoencephalitis demonstrated that CEC-GZL17 infection significantly increases TNF-α expression and inhibits ZO-1 and occludin expressions in brain tissue, indicating that the E coli likely use the mechanism to penetrate the blood-brain barrier via disrupting tight junction architecture, thus leading to the invasion to brain tissue.