Congenital infection with Schistosoma japonicum but not with Schistosoma bovis in sheep

The present study investigated whether Schistosoma japonicum or Schistosoma bois could establish prenatally in lambs. Three ewes were exposed to S. japonicum by intramuscular injection of cercariae, and 3 ewes were exposed to S. bovis cercariae using the leg-emerging technique approximately 2 mo before delivery, and 1 age-matched pregnant ewe served as an uninfected control. The study lasted 18-20 wk after infection, which was 8-9 wk after delivery. All 6 exposed ewes became infected with either S. bovis or S. japonicum. Eight lambs were borne by the 7 ewes, of which 1 (S. bovis exposed) was dead and 1 (S. japonicum exposed) died at delivery. Of the 3 S. japonicum-exposed lambs, 2 were found infected. Four lambs born of S. bovis-exposed ewes were negative. Despite having no worms, these 4 S. bovis-exposed lambs as well as the 1 negative S. japonicum-exposed lamb had, in contrast to the nonexposed control lamb, few, but distinct, liver granulomas dominated by eosinophils and giant cells with large central necrotic areas but with no remnants of eggs or worms. Hence, congenital infection was demonstrated in S. japonicum-infected lambs, but not in S. bovis-infected ones.     

Production of cytokines by monocytes, epithelial and endothelial cells activated by Streptococcus bovis

There are numerous reports documenting the correlation between Streptococcus bovis bacteraemia and endocarditis in conjunction with colonic diseases. The adherence of S. bovis to either buccal or intestinal epithelial cells seems to be the initial process in colonization and subsequent infection of the host, allowing further adhesion of S. bovis to either endothelial cells or extracellular matrix components which leads to infective endocarditis. Bacterial entry at tumour sites is further assisted by the local action of cytokines that promotes vasodilatation and increased capillary permeability. Thus the ability of S. bovis to adhere to and to stimulate human cells may contribute to the pathogenicity of this bacteria. In the present study, we have shown the ability of S. bovis and wall-extracted antigens (WEA) to adhere to human buccal (KB) or intestinal (Caco-2) epithelial cell lines, to human saphenous vein endothelial cells, to human monocytic cell line (THP-1) and to extracellular matrix components (ECM) (fibronectin, collagen and laminin). The fixation of S. bovis on cells was followed by the synthesis of IL-8 from all the cells except Caco-2, whereas S. bovis WEA was able to induce cytokine synthesis from all of them, showing the immunomodulatory effect of S. bovis and S. bovis WEA on different cells.     

Experimental infection of goats with Schistosoma bovis and S. curassoni: comparative pathogenic effects

Specific mortality and morbidity have been quantified in goats experimentally infected with Schistosoma bovis or S. curassoni strains from Niger. The study involved nine animals followed during 380 days after infection with, respectively, 1,800 or 2,400 cercariae. S. bovis was significatively more pathogenic than S. curossoni in terms of mortality, weight loss and packed cell volume decrease. In addition, the intensity of clinical symptoms was significatively and positively correlated to the levels of fecal egg excretion. Compared to non-infected controls, a growth differential of, respectively, 1,600 and 880 grams per month should incite to consider S. bovis and S. curassoni as parasites of serious economical impact in sahelian countries.     

A haemolytic cell-free preparation of Moraxella bovis confers protection against infectious bovine keratoconjunctivitis

Abstract Protection conferred by a cell-free preparation from a haemolytic Moraxella bovis isolate, UQV 148NF, was compared to an equivalent fraction from a non-haemolytic M. bovis isolate, Gordon 26L3, and to a recombinant DNA-derived pili vaccine. Three groups of ten calves were vaccinated twice with one of the three preparations and, together with ten non-vaccinated calves, challenged with virulent M. bovis isolate Dal 2d. Compared to the control group, significant protection was observed in the group receiving the pili vaccine and the group receiving the preparation from haemolytic isolate, UQV 148NF.     

Studies on heterologous resistance between Schistosoma bovis and Fasciola gigantica in Sudanese cattle

Using the local strains of Schistosoma bovis and Fasciola gigantica, it was shown that Sudanese zebu calves with mature primary infections of F. gigantica were highly resistant to challenge with S. bovis cercariae, and vice versa. Liver enzyme tests showed that, in both cases, the primary infections had caused some liver damage. Primary infection with irradiated S. bovis cercariae, which did not cause significant liver damage, did not protect significantly against challenge with F. gigantica.     

Molecular study on cloned endoglucanase gene from rumen bacterium

An endoglucanase gene was subcloned from anaerobic rumen bacterium Ruminococcus flavefaciens strain 17. To express endoglucanase gene in Escherichia coli and Streptococcus bovis JB1, an endoglucanase gene fragment was inserted into pVA838-based shuttle vectors. Removal of endoglucanase gene promoter and expression of endoglucanase by promoter of S. bovis JB1 alpha-amylase gene (pACMCS) was also achieved. Survival of constructs pVACMCI, pTACMC and pACMCS, which carry endoglucanase gene, and stability of endoglucanase gene in S. bovis JB1, were observed. Maximal endoglucanase activities from S. bovis JB1/pVACMCI were 2- to 3-fold higher than from E. coli/pVACMCI. Specific cell activity of E. coli/pACMCS was found to be approximately 2- to -3 fold higher than the both E. coli/pVACMCI and E. coli/pTACMC. Specific cell activity of S. bovis JB1/pACMCS was also found to be approximately 2-fold higher than the both S. bovis/pVACMCI and S. bovis JB1/pTACMC.     

Simple and rapid PCR method for identification of streptococcal species relevant to animal infections based on 23S rDNA sequence

A PCR identification system targeting 23S rDNA sequences for the identification of eight streptococcal species relevant to animal infections (Streptococcus agalactiae, S. bovis, S. canis, S. dysgalactiae, S. equi, S. porcinus, S. suis and S. uberis) was developed. This system consists of two PCR reactions, A and B, in which seven and eight primers, respectively, are used simultaneously, and was designed so that each amplification product indicates a species by its size. A total of 111 cultures, including the type strain of eight species, could be successfully identified and differentiated as individual species, except for the cross reactivity between S. bovis and S. equinus. The developed PCR system can complete the identification procedure for eight streptococcal species through two tube reactions per isolate, and, therefore, might provide a rapid, simple and accurate diagnostic tool for veterinary laboratories.     

Virulent Mycobacterium tuberculosis strains evade apoptosis of infected alveolar macrophages

Human alveolar macrophages (AMphi) undergo apoptosis following infection with Mycobacterium tuberculosis in vitro. Apoptosis of cells infected with intracellular pathogens may benefit the host by eliminating a supportive environment for bacterial growth. The present study compared AMphi apoptosis following infection by M. tuberculosis complex strains of differing virulence and by Mycobacterium kansasii. Avirulent or attenuated bacilli (M. tuberculosis H37Ra, Mycobacterium bovis bacillus Calmette-Guérin, and M. kansasii) induced significantly more AMphi apoptosis than virulent strains (M. tuberculosis H37Rv, Erdman, M. tuberculosis clinical isolate BMC 96.1, and M. bovis wild type). Increased apoptosis was not due to greater intracellular bacterial replication because virulent strains grew more rapidly in AMphi than attenuated strains despite causing less apoptosis. These findings suggest the existence of mycobacterial virulence determinants that modulate the apoptotic response of AMphi to intracellular infection and support the hypothesis that macrophage apoptosis contributes to innate host defense in tuberculosis.     

Molecular survey and genetic identification of Anaplasma species in goats from central and southern China

Anaplasma species are obligate intracellular rickettsial pathogens that impact the health of humans and animals. Few studies have been carried out on Anaplasma infections in central and southern China. This study was conducted to determine the coinfection rates of Anaplasma ovis, A. bovis, and A. phagocytophilum from 262 field blood samples of goats in these regions. The average prevalences of single infection of A. ovis, A. bovis, and A. phagocytophilum were 15.3, 16.0, and 6.1%, respectively. Coinfection of A. ovis and A. bovis was dominant, with an infection rate of 27.1%. Coinfection of A. ovis and A. phagocytophilum was 1.9% and that of A. bovis and A. phagocytophilum was 4.2%. Three-pathogen coinfection was found in three of four investigated provinces with a prevalence between 0 and 5.3%. The accuracy of the PCR results was corroborated by sequencing. Analysis of the 16S rRNA gene sequences of A. bovis and A. phagocytophilum confirmed the presence of these pathogens at the investigated sites and indicated the possible genetic diversity of A. phagocytophilum. Field blood inoculation of experimental animals led to successful identification and observation of the morphological shapes of A. bovis in the infected monocytes of sheep. Phylogenetic study with msp4 sequences of A. ovis indicated that the A. ovis genotypes from sheep in the north differed from the genotypes of goats in the investigated sites.     

Absence of Mycobacterium bovis in feral swine (Sus scrofa) from the southern Texas border region

Free-ranging wildlife, such as feral swine (Sus scrofa), harbor a variety of diseases that are transmissible to livestock and could negatively impact agricultural production. Information is needed regarding the exposure and infection rates of Mycobacterium bovis and many other diseases and parasites in feral swine occurring in the Texas border region. Our main objective was to determine exposure rates and possible infection rates of M. bovis in feral swine by opportunistically sampling animals from the Texas border region. From June to September 2010, we obtained samples from 396 feral swine and tested 98 samples for M. bovis by histopathology and mycobacteriologic culture. We found no evidence of M. bovis infection. We believe that it is important to periodically and strategically sample feral swine for M. bovis in high-risk areas of the United States because they are capable of becoming reservoirs of the disease.     

Characterization of Streptococcus bovis from the rumen of the dromedary camel and Rusa deer

AIMS: Isolation and characterization of Streptococcus bovis from the dromedary camel and Rusa deer. METHODS AND RESULTS: Bacteria were isolated from the rumen contents of four camels and two deer fed lucerne hay by culturing on the semi-selective medium MRS agar. Based on Gram morphology and RFLP analysis seven isolates, MPR1, MPR2, MPR3, MPR4, MPR5, RD09 and RD11 were selected and putatively identified as Streptococcus. The identity of these isolates was later confirmed by comparative DNA sequence analysis of the 16S rRNA gene with the homologous sequence from S. bovis strains, JB1, C14b1, NCFB2476, SbR1, SbR7 and Sb5, from cattle and sheep, and the Streptococcus equinus strain NCD01037T. The percentage similarity amongst all strains was >99%, confirming the identification of the camel isolates as S. bovis. The strains were further characterized by their ability to utilize a range of carbohydrates, the production of volatile fatty acids (VFA) and lactate and the determination of the doubling time in basal medium 10 supplemented with glucose. All the isolates produced l-lactate as a major fermentation end product, while four of five camel isolates produced VFA. The range of carbohydrates utilized by all the strains tested, including those from cattle and sheep were identical, except that all camel isolates and the deer isolate RD11 were additionally able to utilize arabinose. CONCLUSIONS: Streptococcus bovis was successfully isolated from the rumen of camels and deer, and shown by molecular and biochemical characterization to be almost identical to S. bovis isolates from cattle and sheep. SIGNIFICANCE AND IMPACT OF THE STUDY: Streptococcus bovis is considered a key lactic acid producing bacterium from the gastrointestinal tract of ruminants, and has been implicated as a causative agent of lactic acidosis. This study is the first report of the isolation and characterization of S. bovis from the dromedary camel and Rusa deer, and suggests a major contributive role of this bacterium to fermentative acidosis.     

DNA probes distinguish geographical isolates and identify a novel DNA molecule of Babesia bovis

A genomic DNA library of Babesia bovis was screened to identify DNA probe candidates for direct detection of the parasite. Two sequences, Bo6 and Bo25, had the highest sensitivity and further analysis revealed unique characteristics of each of these. Neither sequence hybridized detectably to bovine DNA. Bo6 detected 100 pg of both a Mexican and an Australian isolate of B. bovis, but Bo6 also detected 1.0 ng of Babesia bigemina DNA under identical conditions. A unique characteristic of Bo6 is that it hybridizes to an apparent 7.4-kilobase DNA in undigested genomic DNA of both B. bovis and B. bigemina. The sequence is well conserved between the 2 geographic isolates of B. bovis, but it is apparently divergent in B. bigemina. Bo25 did not hybridize detectably to bovine or B. bigemina DNA. This sequence detected 100 pg of homologous B. bovis Mexican isolate DNA, but the sensitivity was reduced to 1 ng for the Australian isolate DNA. The restriction enzyme profile of the Bo25 sequence in genomic DNA differed markedly in the number, size, and intensity of bands between the 2 B. bovis geographic isolates tested. Thus, the Bo25 sequence can distinguish geographic isolates of B. bovis.     

Bulinus tropicus from Central Kenya acting as a host for Schistosoma bovis

One hundred and twelve snails were collected from two habitats on the Mau Escarpment, Kenya and were provisionally identified as Bulinus tropicus from the characteristics of their shell and soft parts, chromosome number (n = 18), electrophoresis of egg protein on cellulose acetate strip and isoelectric focusing of AcP, GPI, HBDH, MDH and PGM digestive gland enzymes. Of the 55 specimens examined alive in London, 10 were infected with amphistome and schistosome larvae, 9 with amphistome larvae and the remainder were uninfected. The GPI and MDH separations of known infected snails showed two distinct areas of activity: host and parasite. Individual hamsters were exposed to schistosome cercariae emanating from each snail with a double infection (apart from one which died prematurely) and examination of the resulting adult worms showed that all were monomorphic for AcP with a band of enzyme activity at pH 6.45, characteristic of Schistosoma bovis. Examination of eggs found in two infections proved to be S. bovis in shape and size. Exposure of laboratory-bred snails of B. tropicus from the Mau Escarpment and other populations of B. tropicus proved negative. Thus, it is suggested that the presence of the amphistome infection may have a suppressive effect on the immune system of the snail, thereby allowing S. bovis to develop.     

Whole Genome Analysis of Epidemiologically Closely Related Staphylococcus aureus Isolates

The change of the bacteria from colonizers to pathogens is accompanied by a drastic change in expression profiles. These changes may be due to environmental signals or to mutational changes. We therefore compared the whole genome sequences of four sets of S. aureus isolates. Three sets were from the same patients. The isolates of each pair (S1800/S1805, S2396/S2395, S2398/S2397, an isolate from colonization and an isolate from infection, respectively) were obtained within <30 days of each other and the isolate from infection caused skin infections. The isolates were then compared for differences in gene content and SNPs. In addition, a set of isolates from a colonized pig and a farmer from the same farm at the same time (S0462 and S0460) were analyzed. The isolates pair S1800/S1805 showed a difference in a prophage, but these are easily lost or acquired. However, S1805 contained an integrative conjugative element not present in S1800. In addition, 92 SNPs were present in a variety of genes and the isolates S1800 and S1805 were not considered a pair. Between S2395/S2396 two SNPs were present: one was in an intergenic region and one was a synonymous mutation in a putative membrane protein. Between S2397/S2398 only one synonymous mutation in a putative lipoprotein was found. The two farm isolates were very similar and showed 12 SNPs in genes that belong to a number of different functional categories. However, we cannot pinpoint any gene that explains the change from carrier status to infection. The data indicate that differences between the isolate from infection and the colonizing isolate for S2395/S2396 and S2397/S2398 exist as well as between isolates from different hosts, but S1800/S1805 are not clonal.     

Identification and characterization of the proBA operon of Streptococcus bovis.

A genomic DNA library of the rumen bacterium Streptococcus bovis was constructed in Escherichia coli, and recombinant plasmids able to complement proA and proB mutations of the host were found. Southern hybridization and restriction analysis showed that a 3.5-kb fragment of S. bovis DNA contained two genes, organized in an operon and coding for enzymes functionally similar to the glutamyl phosphate reductase-glutamyl kinase enzyme complex that in E. coli catalyzes the first step of proline biosynthesis. Complementation of the E. coli mutations was observed with the fragment inserted in both orientations, which suggested that the S. bovis proBA operon was transcribed from its own promoter. Genetic and biochemical data suggested that the proline biosynthetic pathway of S. bovis is similar to the one previously characterized for E. coli.     

Transmission of Schistosoma bovis in Mkulwe (Mbozi District, Mbeya region, southern highlands of Tanzania)

Various populations of laboratory bred bulinid snails were exposed to miracidia of Schistosoma bovis from Mbozi. The parasite is naturally transmitted by Bulinus globosus in the area. Laboratory infection revealed a good relationship with B. forskalii and B. globosus from Mbozi and a population of B. forskalii from Dar es Salaam (infection rates 100%, 63.6% and 41.7% respectively). Populations of B. globosus and B. nasutus from Dar es Salaam were refractory. It appears that both snail species (B. globosus and B. forskalii) present in Mbozi district transmit S. bovis.