Evaluation of experimental transmission of Candidatus Mycoplasma haemominutum and Mycoplasma haemofelis by Ctenocephalides felis to cats

OBJECTIVE: To determine whether Ctenocephalides felis can transmit Mycoplasma haemofelis (Mhf) and Candidatus Mycoplasma haemominutum (Mhm) through hematophagous activity between cats. ANIMALS: 11 cats. PROCEDURE: 2 cats were carriers of either Mhf or Mhm. Nine cats had negative results via polymerase chain reaction (PCR) assay for Mhf and Mhm DNA; 3 of those cats were infected from the chronic carriers via i.v. inoculation of blood. At the time of maximum organism count for each of the Mycoplasma spp, 1 chamber containing 100 C felis was bandaged to the amplifier cats. Five days later, fleas, feces, larvae, or eggs from each chamber were analyzed for Mycoplasma spp DNA. Viable fleas from the chambers were allocated into new chambers (3 Mhm and 6 Mhf) and attached to na?ve cats for 5 days. Cats were monitored daily for clinical signs and weekly via CBC and PCR assay for infection with Mhf or Mhm for a minimum of 8 weeks. RESULTS: Uptake of Mhf and Mhm DNA into fleas, feces, and, potentially, eggs and larvae was detected. Of the na?ve cats fed on by Mhf-infected fleas, 1 cat transiently yielded positive PCR assay results for Mhf on 1 sampling date without clinical or hematologic changes consistent with Mhf infection. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that hematophagous transfer of Mhm and Mhf into fleas occurred and that C felis is a possible vector for Mhf via hematophagous activity.     

Seroprevalence of Toxoplasma gondii antibodies in clinically ill cats in the United States

OBJECTIVE: To determine regional seroprevalence estimates of Toxoplasma gondi-specific IgM and IgG in clinically ill cats throughout the United States. Sample Population-Sera from 12,628 clinically ill, client-owned cats. PROCEDURE: Toxoplasma gondii-specific IgM and IgG antibodies were detected by use of ELISAs. Sera from clinically ill cats previously submitted for T. gondii antibody testing were sequentially selected from our serum bank and the sample submission paperwork reviewed. The country was divided into 12 geographic regions. Overall prevalence as well as prevalence for each region, age group, season, sex (male vs female), and breed (domestic shorthair vs other) was calculated. Data were analyzed by logistic regression analysis. RESULTS: Overall, 31.6% of the cats were seropositive for T. gondii-specific IgM, IgG, or both. Percentage of cats seropositive for T. gondii antibodies ranged from 16.1% (southwestern United States) to 43.5% (northeastern United States). As age increased, odds of positive T. gondii antibody assay results (IgM alone, IgG alone, and any combination of IgM or IgG) increased. Males were more likely than females to be seropositive for T. gondii antibodies (IgG alone and any combination of IgM or IgG). Domestic shorthair cats were more likely than other breeds to be seropositive for T. gondii antibodies (IgM alone, IgG alone, and any combination of IgM or IgG). CONCLUSIONS AND CLINICAL RELEVANCE: Toxoplasma gondii-specific antibodies are common in serum samples of clinically ill cats from all regions of the United States. Seroprevalence increases as cats age and is higher in male and domestic shorthair cats, compared with females and other breeds.     

The echocardiographic effects of romifidine in dogs with and without prior or concurrent administration of glycopyrrolate

Objective To determine the cardiopulmonary response to romifidine (RO) in the dog with or without prior or concurrent administration of glycopyrrolate. Study Design Randomized, cross‐over experimental study. Animals Six (three male, three female) cross‐bred dogs weighing 23 ± 2.4 kg. Methods Two‐dimensional guided M‐mode echocardiography was performed in conscious dogs simultaneously with measurement of systolic arterial blood pressure (SBP) and heart rate (HR). Dimensions of the left ventricle (LVID), interventricular septum (IVS), and left ventricular free wall (LVFW) were obtained in systole (S) and diastole (D). Amplitude of motion (Amp) of the IVS and LVFW were also measured. From these, measures of wall stress (WS) and fractional shortening (FS) of the left ventricle were derived. Baseline echocardiographic measurements were recorded, following which one of the five treatments was administered. Glycopyrrolate (G) 0.01 mg kg^?1, or saline (S) 0.5 mL, was administered IM as pre‐medication (Gp or Sp), or G was administered concurrently (Gc) with romifidine (RO). Treatments were: T₁, Sp + RO (40 μg kg^?1); T₂, Gp + RO (40 μg kg^?1); T₃, Sp + RO (120 μg kg^?1); T₄, Gp + RO (120 μg kg^?1); and T₅, Sp + Gc +RO (120 μg kg^?1). Romifidine or RO + Gc was administered SC 20 minutes after pre‐medication (time 0), and further measurements were taken 10, 20, 30, 60, and 90 minutes after RO. Results Echocardiographic indices of cardiac systolic function (LVID‐S, FS, Amp‐LVFW) and HR were decreased in RO‐sedated dogs (p < 0.0001) . The magnitude of change in cardiac indices was least with low‐dose RO. At most sampling times, high‐dose RO produced significantly more alteration in cardiac indices. Systolic blood pressure increased in all treatment groups, with the greatest increases in those groups receiving G. Glycopyrrolate significantly increased HR; however, cardiac indices were further reduced. Wall stress significantly increased, with a more dramatic increase in groups receiving G. Conclusions Indices of LV systolic function were reduced in RO‐sedated dogs in a dose‐related manner. Glycopyrrolate further reduced these indices and dramatically increased measurements of wall stress in dogs sedated with RO. Clinical relevance Use of low‐dose RO minimizes cardiac dysfunction; however, it should still be used cautiously in dogs with cardiomyopathy or heart failure. The routine use of G is not recommended to alleviate the bradycardia associated with RO in conscious dogs.     

Epidemiologic study of results of pulsed-field gel electrophoresis of isolates of Rhodococcus equi obtained from horses and horse farms

OBJECTIVE: To compare isolates of Rhodococcus equi on the basis of geographic source and virulence status by use of pulsed-field gel electrophoresis (PFGE). SAMPLE POPULATION: 290 isolates of R equi (218 virulent isolates from foals and 72 avirulent isolates from feces, soil, and respiratory tract samples) obtained between 1985 and 2000 from horses and horse farms from 4 countries. PROCEDURE: DNA from isolates was digested with the restriction enzyme Asel and tested by use of PFGE. Products were analyzed for similarities in banding patterns by use of dendrograms. A similarity matrix was constructed for isolates, and the matrix was tested for nonrandom distributions of similarity values with respect to groupings of interest. RESULTS: There was little grouping of isolates on the basis of country, virulence status, or region within Texas. Isolates of R equi were generally < 80% similar, as determined by use of PFGE. Isolates from the same farm generally were rarely of the same strain. CONCLUSIONS AND CLINICAL RELEVANCE: Considerable chromosomal variability exists among isolates of R equiobtained from the same farm, sites withinTexas, or among countries from various continents. Only rarely will it be possible to link infections to a given site or region on the basis of analysis of isolates by use of PFGE of chromosomal DNA.     

Deletion of Cyp6d4 does not alter toxicity of insecticides to Drosophila melanogaster

Cytochrome P450-dependent monooxygenases are important in the activation and detoxification of numerous insecticides. In this study, a Drosophila melanogaster Cyp6d4 null mutant was used to determine the role of this P450 in insecticide metabolism. This null mutant was generated by imprecise excision of a mobile P element located upstream to the P450 gene Cyp6d4. Comparative analysis between the non-functional mutant and relevant control strains shows that Cyp6d4 does not appear to be involved in the metabolism of chlorfenapyr, cypermethrin, diazinon, imidacloprid, malathion, oxamyl, parathion, or pyrethrum extract, even though these insecticides are known to be activated or detoxified by P450-monooxygenases. No obvious abnormalities in development were seen in the Cyp6d4 null mutant, indicating that Cyp6d4 is not critical for the metabolism of vital endogenous substrates.     

Temporohyoid osteoarthropathy in 33 horses (1993-2000)

A retrospective study of the medical records of 33 horses was performed to determine the clinical and diagnostic abnormalities associated with temporohyoid osteoarthropathy. Data collected from medical records included signalment, presenting complaints, history, physical examination findings, laboratory data, results of diagnostic imaging studies, and treatments. Follow-up information was obtained from a review of case records; by telephone conversation with the owner, veterinarian, or trainer; or by both methods. Of 33 horses with temporohyoid osteoarthropathy, 29 presented with facial nerve (cranial nerve VII) deficits and 23 presented with vestibulocochlear nerve (cranial nerve VIII) deficits. Guttural pouch endoscopy was more reliable than radiography for diagnosis. Of horses with unilateral clinical signs, 22.6% actually had bilateral disease. Magnetic resonance imaging and computed tomography identified the lesions in all horses in which these tests were performed. Of 30 horses for which follow-up information was obtained, 20 (67%) were alive. Eight horses were euthanized and 1 died because of problems associated with temporohyoid osteoarthropathy. Nineteen of 20 surviving horses (95%) were considered by the owner or trainer to be suitable for athletic use. Twelve surviving horses (60%) had residual facial nerve deficits; 11 horses (55%) had residual vestibulocochlear nerve deficits. Horses with temporohyoid osteoarthropathy have a fair prognosis for return to some type of athletic function, but there is risk of acute death. The majority of horses would be expected to have some residual cranial nerve dysfunction, and it could take a year or longer for maximal improvement to occur.     

Strategies for Using eFSH for Superovulating Mares

The standard treatment for superovulation of mares is to administer equine follicle-stimulating hormone (eFSH) for 4 to 5 days to stimulate multiple follicles and human chorionic gonadotropin (hCG) to induce synchronous ovulations. Objectives of this study were: (1) to determine whether a short-term (3-day) eFSH treatment protocol would result in similar ovulation and embryo recovery rates compared with the standard eFSH protocol; (2) to determine the efficacy of a decreasing dose of eFSH (step-down protocol) on ovulation rate and embryo recovery; (3) to compare the efficacy of hCG and recombinant equine luteinizing hormone (reLH) for inducing ovulation in FSH-treated mares; and (4) to compare embryo recovery rates and embryo size when mares are flushed at 6.5 or 7.0 days after ovulation. Forty light-horse mares were used in 2005 (experiment 1) and 20 different mares were used in 2006 (experiment 2). In experiment 1, mares were randomly assigned to one of three treatment groups: (1) untreated controls, (2) standard eFSH treatment (12.5 mg intramuscularly twice daily), and (3) 3-day eFSH treatment. In experiment 2, mares were randomly assigned to one of four treatments: (1) untreated controls, (2) standard eFSH protocol, (3) 3-day eFSH treatment, and (4) step-down eFSH treatment (12.5 mg twice daily day 1, 8.0 mg twice daily day 2, 4.0 mg twice daily day 3). Within each treatment, mares were given either hCG (2,500 IU) or equine LH (750 mg, EquiPure LH; reLH) to induce synchronized ovulations. Embryo recovery was performed either 6.5 or 7.0 days after ovulation. In experiment 1, numbers of preovulatory follicles and ovulations were less for mares in the 3-day treatment group than the standard group, but were greater than for controls. Embryo recovery per flush was higher in the standard group (2.6) than the 3-day eFSH treatment (0.8) or control groups (0.8). In experiment 2, the number of preovulatory follicles and number of ovulations were greater in the standard and 3-day treatment groups than in control and step-down groups. The percent embryo recovery per ovulation and mean embryo grade were similar for all groups; however, the embryo recovery per flush was higher for mares in the standard treatment than controls (1.3 vs 0.6) but was similar to the 3-day (1.1) and step-down (0.8) treatments. Embryo recovery was similar for flushes performed on days 6.5 and 7.0 post-ovulation. The percentage of control mares ovulating within 48 hours in response to hCG or reLH was similar. In contrast, a higher percentage of eFSH-treated mares ovulated within 48 hours in response to reLH than hCG (92% vs 71%). In both years, the 3-day eFSH treatment protocol resulted in a greater number of preovulatory follicles and a greater number of ovulations than untreated controls. Unfortunately, the increased ovulation rate for mares administered eFSH for 3 days did not result in a greater number of embryos recovered per flush in either year. Use of a step-down eFSH treatment protocol resulted in fewer preovulatory follicles, fewer ovulations, and fewer embryos as compared with the standard eFSH treatment. In conclusion, the standard eFSH treatment resulted in a greater embryo recovery rate per cycle than either the 3-day or step-down treatment protocols. Recombinant equine LH was more effective than hCG in causing ovulation in eFSH-treated mares.     

The ecology of horse cyathostomin infective larvae (Nematoda-Cyathostominae) in tropical southeast Brazil

Experimental studies about the recovery, survival and migration to pasture of cyathostomin infective larvae (L(3)) from fresh feces depositions were conducted from February 2005 to March 2007 in a tropical region of southeast Brazil. Grass and feces were collected weekly at 8 a.m., 1 and 5 p.m. and processed by the Baermann technique. Multivariate analysis (principal components method) showed the influence of time and environmental variables on the number of infective larvae recovered from the feces and pasture. In the rainy period (October-March), more infective larvae were recovered on the feces and grass apex. In contrast, in the dry period (April-September), the recovery was higher only on the grass base, as well as the L(3) survival on feces and grass. More larvae were recovered at 8 a.m., except from the grass apex, where the highest recovery was at 1 p.m. Few studies investigating the seasonal transmission of equine cyathostomin have been conducted in South American tropical climates. These results demonstrate that in tropical conditions L(3) are available on feces and pasture throughout the year. Knowledge of climatic influences on the development and survival of L(3) is crucial to designing integrated parasite control programs that provide effective protection while slowing the development of anthelmintic resistance.     

A possible role for Clostridium difficile in the etiology of calf enteritis

Clostridium difficile was investigated as a possible cause of enteritis in calves. The organism and its toxins (TcdA and TcdB), respectively, were found in 25.3% and 22.9% of stool samples from diarrheic calves. Culture positive samples were more likely than culture negative samples to be toxin positive. However, toxin positive stools were more common among nondiarrheic calves, but diarrheic calves were nearly twice as likely to be culture positive. Ribotype 078 was dominant among isolates. Salmonella sp. was isolated from both diarrheic and nondiarrheic calves, but large numbers of E. coli were found more commonly in diarrheic calves than in nondiarrheic animals. Prevalence rates for coronavirus and Cryptosporidium sp. were substantially higher in nondiarrheic calves than in diarrheic, but rates of detection of rotavirus and Giardia sp. were more nearly equal between groups. Lesions in naturally infected calves included superficial mucosal erosion with associated fibrinous exudates. Neutrophils and eosinophils infiltrated lamina propria. Large Gram-positive rods morphologically compatible with C. difficile were abundant in the colonic lumen and the organism was isolated by bacteriologic culture. Toxins were found throughout the colon. Purified toxins A and B (individually and conjointly) caused comparable lesions, as well as fluid accumulation, in ligated intestinal loops. Our findings are in substantial agreement with those of others [Rodriguez-Palacios, A., Stampfli, H.R., Duffield, T., Peregrine, A.S., Trotz-Williams, L.A., Arroyo, L.G., Brazier, J.S., Weese, J.S., 2006. Clostridium difficile PCR ribotypes in calves, Canada. Emerg. Infect. Dis. 12, 1730-1736; Porter, M.C., Reggiardo, C., Bueschel, D.M., Keel, M.K., Songer, J.G., 2002. Association of Clostridium difficile with bovine neonatal diarrhea. Proc. 45th Ann. Mtg. Amer. Assoc. Vet. Lab. Diagn., St. Louis, MO, U.S.A.] and add strength to a working hypothesis that C. difficile infection and the accompanying intoxication can manifest as diarrhea in calves. It seems clear that calves serve as multiplying hosts for this organism.