Cholera remains a significant cause of mortality in developing countries. Outbreaks of the disease are associated with poverty, lack of potable water and poor sanitation. The survival and persistence of Vibrio cholerae in water has been shown to depend on physico-chemical factors. We studied water sources in Bepanda, an overcrowded neighbourhood in Douala, Cameroon, with limited access to portable water and very poor sanitary conditions as reservoirs of V. cholerae. We analysed 318 samples from various sources (well, tap, stream) from February to July 2009 using standard microbiological techniques and characterised isolates serologically using the polyvalent O1/O139 antisera. Susceptibility to antibiotics previously used for cholera treatment in Douala was studied using the disk diffusion method. Physico-chemical factors (temperature, pH and salinity) that could maintain the endemicity of the organism were analysed using standard methods. Eighty-seven (27.4%) samples were contaminated, with high isolation rates being obtained from streams (52.4%) and wells (29.8%). The number of isolates was significantly higher (P < 0.05) in the rainy season (35.5%). We detected 23 (24%) O1 serogroup isolates in streams and wells, whilst 64 (66.6%) were non-O1/non-O139. Temperature and salinity correlated positively with the occurrence of the organisms. All isolates were susceptible to fluoroquinolones but high resistance rates to trimethoprim or sulfamethozaxole and tetracycline were observed. Vibrio cholerae is endemic in Bepanda with O1 and non-O1/non-O139 serogroups co-existing in the streams and wells hence the possibility of future outbreaks of cholera if sanitation and drinking water quality are not improved. Temperature and salinity are amongst the factors maintaining the endemicity of the organism.

Foot-and-mouth disease (FMD) is endemic in Tanzania. Since the first reports in 1954, FMD has caused significant economic losses in the country due to mortality and morbidity of livestock and costs associated with controlling the disease. The aim of this study was to review the serotype and genetic relationships of the FMD virus (FMDV) recovered from outbreaks in Tanzania, and compare them with viruses detected from elsewhere in the sub-Saharan region. At the World Reference Laboratory for foot-and-mouth disease (WRLFMD), a total of 106 FMD viruses have been isolated from samples collected between 1967 and 2009 from northern, southern, eastern and central parts of Tanzania. The presence of FMDV was determined by laboratory methods such as VI, CF, antigen ELISA and RT-PCR. Phylogenies of VP1 sequences were determined by the Neighbour-joining method. Foot-and-mouth disease virus SAT1 was the most frequent serotype (46.2%; n = 49) isolated in Tanzania followed by O (26.4%; n = 27), A (14.1%; n = 15) and SAT 2 (11.3%; n = 13). Genotyping showed that type O viruses fell into either the EAST AFRICA 1 (EA-1) or EA-2 topotypes, type A’s into the AFRICA topotype (genotype I), type SAT 1’s into topotype I and type SAT 2’s into topotype IV. This study reveals that serotypes A, O, SAT1 and SAT2 cause FMD outbreaks in Tanzania. Recent samples from outbreaks in 2008, 2009 and 2010 have been typed as serotypes A, O, SAT1 and SAT2. Phylogenetic analysis of FMDV isolates from Tanzania showed that they are genetically related to lineages and topotypes from West and East Africa. In Tanzania, lack of comprehensive animal movement records and inconsistent vaccination programs make it difficult to determine the exact source of FMD outbreaks or to trace the transmission of the disease over time. Therefore, further collection and analysis of samples from domestic and wild animals, together with improved local epidemiological investigation of FMD outbreaks is required to elucidate the complex epidemiology of FMD in the sub-Saharan region.

Objective: To assess the agreement and reliability of cardiac measurements obtained with 3 echocardiographic techniques in anesthetized red-tailed hawks (Buteo jamaicensis). Animals: 10 red-tailed hawks. Procedures: Transcoelomic, contrast transcoelomic, and transesophageal echocardiographic evaluations of the hawks were performed, and cineloops of imaging planes were recorded. Three observers performed echocardiographic measurements of cardiac variables 3 times on 3 days. The order in which hawks were assessed and echocardiographic techniques were used was randomized. Results were analyzed with linear mixed modeling, agreement was assessed with intraclass correlation coefficients, and variation was estimated with coefficients of variation. Results: Significant differences were evident among the 3 echocardiographic methods for most measurements, and the agreement among findings was generally low. Interobserver agreement was generally low to medium. Intraobserver agreement was generally medium to high. Overall, better agreement was achieved for the left ventricular measurements and for the transesophageal approach than for other measurements and techniques. Conclusions and Clinical Relevance: Echocardiographic measurements in hawks were not reliable, except when the left ventricle was measured by the same observer. Furthermore, cardiac morphometric measurements may not be clinically important. When measurements are required, one needs to consider that follow-up measurements should be performed by the same echocardiographer and should show at least a 20% difference from initial measurements to be confident that any difference is genuine.

Objective: To investigate the relationship between runt-related transcription factor 2 (RUNX2) expression in canine nucleus pulposus (NP) cells and intervertebral disk aging in chondrodystrophoid dogs. Animals: 7 healthy Beagles (mean age, 35.6 months) and 11 Dachshunds with herniated disks (mean age, 61 months). Procedures: All dogs underwent MRI examination of the thoracic and lumbar vertebral column immediately before sample collection under general anesthesia. The disk center–to–CSF T2-weighted signal intensity ratio was determined for healthy Beagles. Samples of NP were obtained from nonherniated disks in healthy Beagles and from herniated disks during surgical treatment of hospitalized Dachshunds. Samples were evaluated for RUNX2 and matrix metalloproteinase 13 transcript expression via reverse transcriptase PCR assay; RUNX2 protein expression was evaluated via immunohistochemical analysis, and correlation between these variables and age of dogs was evaluated. A 3′ and 5′ rapid amplification of cDNA ends method was used to identify the RUNX2 coding region. Results: RUNX2 cDNA had > 97% conservation with the human cDNA sequence and approximately 95% conservation with the mouse cDNA sequence; RUNX2 and matrix metalloproteinase 13 mRNA expression and RUNX2 protein expression in NP cells were positively correlated with age. The disk center–to–CSF T2-weighted signal intensity ratio was negatively correlated with RUNX2 protein expression in the NP of healthy dogs. Conclusions and Clinical Relevance: Results indicated that RUNX2 mRNA and protein expression in the NP are enhanced in aging intervertebral disks in dogs.

Objective: To develop in genetically engineered mice an alternative screening method for evaluation of P-glycoprotein substrate toxicosis in ivermectin-sensitive Collies. Animals: 14 wild-type C57BL/6J mice (controls) and 21 genetically engineered mice in which the abcb1a and abcb1b genes were disrupted and the mutated canine ABCB1 gene was inserted. Procedures: Mice were allocated to receive 10 mg of ivermectin/kg via SC injection (n = 30) or a vehicle-only formulation of propylene glycol and glycerol formal (5). Each was observed for clinical signs of toxic effects from 0 to 7 hours following drug administration. Results: After ivermectin administration, considerable differences were observed in drug sensitivity between the 2 types of mice. The genetically engineered mice with the mutated canine ABCB1 gene had signs of severe sensitivity to ivermectin, characterized by progressive lethargy, ataxia, and tremors, whereas the wild-type control mice developed no remarkable effects related to the ivermectin. Conclusions and Clinical Relevance: The ivermectin sensitivity modeled in the transgenic mice closely resembled the lethargy, stupor, disorientation, and loss of coordination observed in ivermectin-sensitive Collies with the ABCB1–1Δ mutation. As such, the model has the potential to facilitate toxicity assessments of certain drugs for dogs that are P-glycoprotein substrates, and it may serve to reduce the use of dogs in avermectin derivative safety studies that are part of the new animal drug approval process.

Objective: To evaluate the anterior chamber approach and energy levels for endoscopic cyclophotocoagulation (ECPC) and assess ECPC-induced tissue damage in phakic eyes of bovine cadavers. Sample: 12 bovine cadaver eyes. Procedures: Angle of reach was measured in 6 eyes following placement of a curved endoscopic probe through multiple corneal incisions. In another 6 eyes, each ocular quadrant underwent ECPC at 1 of 3 energy levels (0.75, 0.90, and 1.05 J) or remained untreated. Visible effects on tissues (whitening and contraction of ciliary processes) were scored (scale of 0 [no effects] to 6 [severe effects]), and severity and extent of histologic damage to the pigmented and nonpigmented ciliary epithelium and fibromuscular stroma were each scored (scale of 0 [no effect] to 3 [severe effect]) and summed for each quadrant. Overall mean scores for 6 quadrants/treatment were calculated. Results: Mean ± SD combined angle of reach was 148 ± 24° (range, 123 ± 23° [ventromedial] to 174 ± 11° [dorsolateral]). At the 0.75-, 0.90-, and 1.05-J levels, mean visible tissue effect scores were 3.12 ± 0.47, 3.86 ± 0.35, and 4.68 ± 0.58, respectively; mean histologic damage scores were 4.79 ± 1.38 (mild damage), 6.82 ± 1.47 (moderate damage), and 9.37 ± 1.42 (severe damage), respectively. Occasional popping noises (venting of vaporized interstitial water) were heard at the 1.05-J level. Conclusions and Clinical Relevance: Multiple incisions were necessary to facilitate 360° ECPC treatment in bovine eyes. For ECPC in vivo, the 0.75- and 0.90-J energy levels had the potential to effectively treat the ciliary epithelium.

This study investigated and compared the antimicrobial resistance patterns and ribotypes of Staphylococcus aureus isolated from pig tonsils and cow’s milk in China. A total of 90 isolates of S. aureus was included: 42 strains were isolated from tonsils of pigs and 48 from half-udder milk. The broth microdilution method and the double-disc diffusion test (D test) were used for antimicrobial susceptibility testing. The mecA gene for methicillin-resistant S. aureus (MRSA) and the ermA, ermB, ermC, and msrA genes for erythromycin-resistant strains were detected by polymerase chain reaction (PCR). The isolates were ribotyped with the Riboprinter system. The highest frequency of resistance was observed with clindamycin (91.1%), followed by penicillin (90.0%), and erythromycin (85.6%). All strains were susceptible to vancomycin and trimethoprim-sulfamethoxazole. The D test showed that 54.5% (42/77) of erythromycin-resistant isolates had the constitutive resistance phenotype and 45.5% (35/77) had the inducible resistance phenotype to clindamycin. A higher proportion of resistance to cephalosporins, macrolides, fluoroquinolones, and pleuromutilins was observed in pig isolates than in milk isolates (P < 0.05). The mecA gene was detected in all MRSA isolates; 89.6% of erythromycin-resistant strains harbored the ermC gene and 16.9% harbored the ermB gene. A total of 35 different ribogroups was found among the isolates investigated; 83.3% of pig strains belonged to 1 cluster with a similarity coefficient of 0.84. In contrast, 3 main clusters were observed among 68.8% of milk strains, which indicates a high degree of host specificity.

Objective: To determine the efficacy of an avirulent Lawsonia intracellularis vaccine in preventing proliferative enteropathy in weanling foals. Animals: 12 healthy weanling foals. Procedures: Foals were randomly assigned to a vaccinated, nonvaccinated, or control group. Vaccinated foals received an avirulent porcine L intracellularis frozen-thawed vaccine intrarectally 60 and 30 days prior to experimental challenge. On day 1, vaccinated and nonvaccinated foals were challenged via nasogastric intubation with a virulent heterologous isolate of L intracellularis. Control foals were not challenged. Clinical observation and ultrasonographic evaluation of the small intestine were performed, and body weight, serum concentration of total solids, fecal excretion of L intracellularis, and seroconversion were measured for each foal until day 56. Diseased foals were treated with antimicrobials and supportive are. Results: None of the 4 vaccinated foals developed clinical disease following challenge with virulent L intracellularis. Three of 4 nonvaccinated foals developed moderate to severe clinical signs compatible with proliferative enteropathy, hypoproteinemia, and thickened small intestinal loops. Vaccinated foals had significantly less fecal shedding of L intracellularis than nonvaccinated foals. Serologic responses between vaccinated and nonvaccinated foals after challenge were similar. Control foals remained clinically unaffected with no evidence of fecal shedding and seroconversion. Conclusions and Clinical Relevance: Intrarectal administration of a commercial avirulent porcine vaccine against L intracellularis resulted in complete protection against proliferative enteropathy in the foals in this study and may also reduce environmental contamination with the organism on endemic farms.

Objective: To establish practical doses and administration frequencies of fondaparinux for cats that would approximate human therapeutic peak and trough plasma anti–factor Xa activities for thromboprophylaxis (TP) and thrombosis treatment (TT) protocols. Animals: 6 healthy adult purpose-bred cats. Procedures: Dosage protocols for TP and TT were selected on the basis of a single compartment pharmacokinetic model incorporating data from humans but modified to account for the higher body weight–normalized cardiac output of cats. Fondaparinux was administered at 0.06 mg/kg, SC, every 12 hours (TP) for 7 days in one session, and 0.20 mg/kg, SC, every 12 hours (TT) for 7 days in another, with a minimum of 1 week separating the sessions. Plasma anti–factor Xa activity was measured before fondaparinux administration (day 1) and at 2 (peak) and 12 (trough) hours after drug administration on days 1 and 7. Platelet aggregation and thromobelastographic (TEG) parameters were also measured 2 hours after drug administration on day 7. Results: Peak plasma anti–factor Xa activities on day 7 for TP (median, 0.59 mg/L; range, 0.36 to 0.77 mg/L) and TT (median, 1.66 mg/L; range, 1.52 to 2.00 mg/L) protocols were within therapeutic ranges for humans. However, only the TP protocol achieved trough anti–factor Xa activity considered therapeutic in humans (median, 0.19 mg/L; range, 0.00 to 0.37 mg/L) on day 7. There were significant changes in the TEG parameters at peak for the TT protocol, suggesting a hypocoagulable state. No significant changes in platelet aggregation were evident for either protocol. Conclusions and Clinical Relevance: A fondaparinux dosage of 0.06 or 0.20 mg/kg, SC, every 12 hours, was sufficient to achieve a peak plasma anti–factor Xa activity in cats that has been deemed therapeutic in humans. This study provided preliminary data necessary to perform fondaparinux dose-determination and clinical efficacy studies.

Objective: To develop and evaluate a rapid and accurate assay involving PCR amplification and electrospray ionization mass spectrometry of nucleic acid extracts from whole blood samples for the detection of Dirofilaria immitis infection in dogs. Sample: Whole blood nucleic acid extracts from 29 dogs experimentally infected with D immitis (and in which circulating D immitis antigen was detected) and 10 uninfected dogs. Procedures: 16 of the 29 whole blood samples from infected dogs were examined at the time of collection for circulating microfilaria. Nucleic acids were extracted from all whole blood specimens and underwent PCR amplification with 12 PCR primer pairs designed to detect a wide range of pathogens (including the Wolbachia endosymbiont of D immitis) and electrospray ionization mass spectrometry. Results: On the basis of assay results, heartworm infection was detected in 13 of 13 antigen-positive dogs of unknown microfilaria status, 11 of 11 antigen-positive dogs with circulating microfilaria, 0 of 3 antigen-positive dogs tested at 3 months after larval infection, 0 of 2 antigen-positive dogs with occult infections, and 0 of 10 uninfected dogs. Conclusions and Clinical Relevance: With the assay under investigation, it was possible to identify D immitis infection in dogs with circulating microfilaria via detection of the obligate Wolbachia endosymbiont of D immitis. It was not possible to identify dogs with occult infections, which suggested that circulating microfilaria must be present to detect infection with this assay, although further studies would be required to verify that finding.