Characterization of a wild-type strain of Francisella tularensis isolated from a cat.

Francisella tularensis type A is the primary cause of tularemia in animals and humans in North America. The majority of research on F. tularensis has been done with the attenuated live vaccine strain (LVS), which is a type B, but very few wild-type F. tularensis strains have been characterized. A gram-negative coccobacillus that was isolated in pure culture from the lungs of a cat that died after being lost for 5 days was received for identification at the Virginia-Maryland Regional College of Veterinary Medicine Teaching hospital. The isolate (strain TI0902) was not identified (or was misidentified) by commercial identification systems; however, it was identified as F. tularensis subspecies tularensis (type A) by sequencing a portion of the 16S ribosomal RNA gene. Furthermore, repetitive extragenic palindromic sequences-polymerase chain reaction amplified a 4-kb DNA fragment from TI0902 that was characteristic of F. tularensis type A but not type B. The electrophoretic profile of the lipopolysaccharide of strain TI0902 was identical to that of the LVS by Western blotting with antiserum to LVS. The protein-enriched outer membrane of strain TI0902 contained 6-8 proteins, which were similar in molecular size to those from the LVS. Electron microscopy of negatively stained and alcian blue-stained LVS and TI0902 cells showed that both strains were coccobacillary in shape and may be encapsulated. However, after mouse challenge, the TI0902 strain was clearly more virulent than the LVS strain. Results of this study indicate that the genotype and phenotype of wild-type F. tularensis type A strain TI0902 is similar, but not identical, to that of the LVS strain. Further studies will help determine whether pathogenesis and host-pathogen interactions are also similar between the 2 strains.     

Prevalence and risk factors associated with bovine brucellosis in herds under extensive production system in southwestern Nigeria

Using a cross-sectional survey, we determined the prevalence and risk factors associated with bovine brucellosis in herds under extensive production system in southwestern Nigeria. Antibodies to Brucella species in serum samples were tested using the Rose Bengal test (RBT) and competitive enzyme-linked immunosorbent assay (cELISA); for milk, the milk ring test (MRT) and indirect-ELISA (i-ELISA) were used. Questionnaire was administered to cattle herdsmen to determine factors predisposing the animals to bovine brucellosis. Data were analyzed using STATA 12. From 513 serum and 635 milk samples tested among 120 herds, overall animal-level prevalence of 10.1% (95% CI 7.5–12.7%) and 20.2% (95% CI 17.1–23.3%) were recorded by RBT and MRT, respectively; while 9.4% (95% CI 6.9–11.9%) and 17.8% (95% CI 14.8–20.8%) were obtained using cELISA and i-ELISA, respectively. In all, from the 120 herds tested, 29.2% and 43.3% were positive by RBT and MRT, respectively. Multivariable logistic regression revealed that herd location (OR?=?8.12, 95% CI 1.68–38.90) and improper disposal of placenta/fetus (OR?=?17.33, 95% CI 4.81–62.33) were predictors for a seropositive herd using RBT; while herd location (OR?=?5.13, 95% CI 1.27–20.28), large herd size (OR?=?2.62, 95% CI 1.15–5.85), and occurrence of abortion for a year or more (OR?=?4.62, 95% CI 1.53–13.71) were predictors of seropositivity to antibodies to Brucella spp. using MRT. We found high prevalence of brucellosis in cattle herds under extensive management system in southwestern Nigeria. Urgent and coordinated control strategies are required to mitigate this problem.

     

Concepts and Direction of Induced Systemic Resistance in Plants and its Application

Resistance to plant disease is often specific and metabolites and receptors contributing to this specificity may have specific structures. However, simple, structurally-unrelated compounds induce systemic resistance in unrelated plants to diverse pathogens including fungi, bacteria and viruses. Both resistance and induced systemic resistance (ISR) are associated with the rapid accumulation of the same structurally unrelated putative defense compounds that have diverse functions. It has been suggested that cultivar (race)-specific resistance is initiated by the specific interaction of a pathogen product (or pathogen induced product) and a plant receptor. However, restricted infection by pathogens can result in ISR and many different compounds can cause ISR. It is thus evident that there are both specific and non-specific routes to the master switch for ISR and there may be more than one master switch. Are reactive oxygen species and free radicals regulating the master switch(es) via both routes? It is also evident there are many switches, other than the master switch. Adding to the complexity of resistance and ISR are the observations that different compounds and pathways may mediate different biochemical resistances. Activation of one of the pathways may antagonize or enhance the activation or effectiveness of another. The review will address these complexities and questions and propose directions of research which require high priority. Factors which encourage and suppress the application of ISR in agriculture will also be addressed.     

Pharmacokinetic comparison of six anthelmintics in sheep,goats, and cattle

This study was initiated to determine whether a comparative pharmacokinetic (PK) approach could be used to expand the pool of approved anthelmintics for minor ruminant species. Accordingly, the PK profiles of six anthelmintics (levamisole, albendazole, fenbendazole, moxidectin, doramectin, and ivermectin) in sheep, goats, and cattle were determined. The PK values determined for each anthelmintic included T_max, T_last, C_max, AUC, AUC/dose, and C_max/dose. The results of this study demonstrate that a comparative PK approach does not show commonality in the way these six anthelmintics are individually processed by these three ruminants. While some drugs demonstrated identical PK profiles between sheep and goats, none of these drugs demonstrated PK profiles in sheep and goats comparable to the PK profiles found in cattle. The results from this study suggest drug approval across these three ruminants is not a viable concept. However, the resulting PK profiles for each combination of drug and ruminant species represents a new dataset that can be used to support the US FDA Center for Veterinary Medicine's Minor Use/Minor Species indexing process for drug approvals in minor species such as sheep and goats.     

Pulsed-field gel electrophoresis patterns and antimicrobial susceptibility phenotypes for coagulase-positive staphylococcal isolates from pustules and carriage sites in dogs with superficial bacterial folliculitis

OBJECTIVE: To determine whether coagulase-positive staphylococcal isolates that are genotypically the same strain obtained from pustules and carriage sites of individual dogs with superficial bacterial folliculitis have the same antimicrobial susceptibility phenotype. ANIMALS: 40 dogs with superficial bacterial folliculitis. PROCEDURES: Samples were obtained from 3 pustules and 3 carriage sites (ie, anus, nonlesional axillary skin, and nasal mucosa) for bacterial culture, morphologic identification, Gram staining, catalase and coagulase testing, antimicrobial susceptibility testing, speciation, and pulsed-field gel electrophoresis (PFGE). RESULTS: 223 isolates from pustules and carriage sites were included. Seventeen susceptibility phenotypes were found among isolates. One hundred twenty-eight (100%) isolates from pustules and 95 (100%) isolates from carriage sites were susceptible to cephalothin; 128 (100%) isolates from pustules and 94 (98.9%) isolates from carriage sites were susceptible to amoxicillin-clavulanic acid; 114 (89.1%) isolates from pustules and 82 (86.3%) isolates from carriage sites were susceptible to erythromycin and lincomycin hydrochloride; and 103 (80.5%) isolates from pustules and 70 (73.7%) isolates from carriage sites were susceptible to trimethoprim-sulfamethoxazole. In 37 of 39 (94.9%) dogs, isolates with the same PFGE pattern from multiple pustules had the same susceptibility phenotype. In 21 of 33 (63.6%) dogs, isolates from multiple carriage sites with the same PFGE pattern had the same susceptibility phenotype. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs with superficial bacterial folliculitis, most coagulase-positive staphylococcal isolates from pustules that are genotypically the same strain will have the same susceptibility phenotype and treatment may be based on empiric antimicrobial selection or susceptibility testing of 1 lesional isolate.