Power and robustness of three whole genome association mapping approaches in selected populations

Selection is known to influence the linkage disequilibrium (LD) pattern in livestock populations. Spurious LD may lead to a higher number of false-positive signals in whole genome association mapping experiments. We compared three approaches for whole genome association mapping in a simulation study: single marker regression (SMR), a two-step approach, which analyses residuals corrected for family effects with an SMR (GRAMMAR), and a combined linkage and LD approach, which applies the quantitative transmission disequilibrium test to the Mendelian sampling term (MTDT). Three different scenarios were simulated: idealized random mating, limited number of parents and directional selection. The number of false-positive associations increased when the number of parents was limited. Mapping accuracy was the worst in the scenario with directional selection for all approaches. As SMR produced a high number of false-positive signals in small populations, results of whole genome scans in livestock analysed with SMR should be considered with caution. GRAMMAR was the most accurate approach, but also the least powerful one. The Bonferroni-corrected significance threshold seemed to be too stringent for this approach. Results obtained with MTDT changed only slightly with selected populations. MTDT combined sufficient power with a manageable number of false-positive associations in all scenarios.     

Candidate gene expression and intramuscular fat content in pigs

Seventy-two pigs of three genetic groups (Brazilian indigenous breed Piau, Commercial line and Crossbred) of both sexes were slaughtered at four live weights (30, 60, 90 and 120 kg). Intramuscular fat (IMF) content in Longissimus dorsi muscle of each animal was extracted and correlated with candidate gene mRNA expression (ATN1, EEF1A2, FABP3, LDLR, MGP, OBSCN, PDHB, TRDN and RYR1). Within slaughter weight of 120 kg, Piau and Crossbred pigs showed higher IMF content (p < 0.05) than commercial animals, with 2.48, 2.08 and 1.00% respectively. Barrows presented higher values of IMF (p < 0.05) than gilts (1.54 and 1.30% respectively). Gene expression of EEF1A2, FABP3, LDLR, OBSCN, PDHB, TRDN and RYR1 were correlated with IMF (p < 0.05) using the whole dataset. For Piau data only, expression of FABP3, LDLR, MGP, OBSCN, PDHB, TRDN and RYR1 showed correlation with IMF (p < 0.05). Genes that have important roles in lipid transportation inside the cell (FABP3) and tissues (LDLR) showed correlation with IMF of, respectively, 0.68 and 0.63 using the whole data set, and 0.90 and 0.91 using data from Piau animals. The highly positive correlation of the LDLR and FAPB3 expression with IMF content may confirm that these genes are important for fat deposition in the porcine L. dorsi muscle.     

Experimental infection of dogs (Canis familiaris) with sporulated oocysts of Neospora caninum

Neospora caninum is widely distributed in the world and this parasite is one of the major causes of abortion in cattle. Dogs and coyotes are definitive hosts of N. caninum and several species of domestic and wild animals are intermediate hosts. Dogs can become infected by the ingestion of tissues containing cysts and then excrete oocysts. It is not yet known whether sporulated oocysts are able to induce a patent infection in dogs, i.e. a shedding of N. caninum oocysts in feces. The objective of this study was to experimentally examine the infection of dogs by sporulated oocysts. The oocysts used in the experiment were obtained by feeding dogs with brain of buffaloes (Bubalus bubalis) positive for anti-N. caninum antibodies by indirect fluorescent antibody test (IFAT ≥200). Oocysts shed by these dogs were confirmed to be N. caninum by molecular methods and by bioassay in gerbils, and sporulated N. caninum oocysts were used for the oral infection of four dogs. The dogs were 8 weeks old and negative for antibodies to N. caninum and Toxoplasma gondii. Dogs 1 and 4 received an inoculum of 10,000 sporulated oocysts each; dog 2 an inoculum of 5000 sporulated oocysts and dog 3 received 1000 sporulated oocysts of N. caninum. The total feces excreted by these dogs were collected and examined daily for a period of 30 days. No oocysts were found in their feces. The dogs were monitored monthly for a 6-month period to observe a possible seroconversion and when this occurred the animals were eliminated from the experiment. Dogs 1 and 4 seroconverted 1 month after the infection with titer, in the IFAT, of 1600 and 800, respectively; the other two dogs presented no seroconvertion during the 6-month period. Dogs 1 and 2 were euthanized 180 days after infection and were examined for the detection of N. caninum in tissues (brain, muscle, lymph node, liver, lung, heart and bone marrow) by immunohistochemistry and PCR with negative results in both techniques. Bioassay in gerbils with brain of these dogs was also performed and again the results were negative. In conclusion, dogs infected with sporulated oocysts of N. caninum were not able to shed oocysts in feces. However, a higher dose of infection stimulated the production of antibodies against N. caninum in the dogs.     

High mortality and lesions of the central nervous system in trypanosomosis by Trypanosoma vivax in Brazilian hair sheep

Here, we report an outbreak of Trypanosoma vivax-induced trypanosomosis in Brazilian hair sheep on a farm in Paraíba state, a non-endemic region in northeastern Brazilian. Of 306 total sheep, 240 showed clinical signs and 216 died. Clinical signs included anorexia, lethargy, anemia, rough hair coat, weight loss, submandibular edema, abortion, and in some cases, neurological signs such as head pressing, lateral recumbence, paddling movements and muscle tremors. T. vivax was identified by blood smear analysis and polymerase chain reaction (PCR). At necropsy, animals exhibited watery blood, pale tissue coloring, and the presence of liquid in the peritoneal cavity and pericardial sac. Histologically, nonsuppurative myocarditis and meningoencephalitis with areas of malacia were observed. After treatment, no parasites were detected by blood smear analysis or PCR. Cattle and buffalo that remained in the same pasture were also infected but presented with asymptomatic infections. Epidemiological data suggest that T. vivax was introduced to the farm and the susceptible flock by buffalos that were asymptomatic carriers of the infection; T. vivax was most likely transmitted by Tabanus spp. bites and also iatrogenically.     

Leishmania spp. in Didelphis albiventris and Micoureus paraguayanus (Didelphimorphia: Didelphidae) of Brazil

Leishmaniasis is kept in nature by the participation of several animal species. This study evaluated the presence of Leishmania spp. in skin samples of free-ranging marsupials Micoureus paraguayanus (n=95) and Didelphis albiventris (n=191), captured in Morro do Diabo State Park and in sections of its surrounding forest, in the region of Pontal do Paranapanema, S?o Paulo State, Brazil. The samples were tested for the presence of kDNA of Leishmania spp. by polymerase chain reaction (PCR) and by real time PCR (qPCR). All samples from D. albiventris tested by PCR were negative for the presence of kDNA of Leishmania spp. However, when tested by qPCR, the positivity was 1.6%. A positivity of 7.4% by PCR and 11.6% by qPCR was observed for M. paraguayanus. Sixty-four per cent (9/14) of positive animals were limited to the same forest fragment. Presence of Leishmania (Leishmania) amazonensis and Leishmania (Viannia) braziliensis was detected in M. paraguayanus samples. While D. albiventris is the most studied marsupial species due to its urban habits, other marsupial species such as M. paraguayanus can be potential reservoirs of Leishmania spp. and should also be studied.     

Use of crude, FML and rK39 antigens in ELISA to detect anti-Leishmania spp. antibodies in Felis catus

Visceral leishmaniasis is a disease caused by Leishmania (Leishmania) chagasi and represents a serious public health problem. The dog is the main urban reservoir of the disease; however, investigations regarding the occurrence and epidemiological importance of leishmaniasis in cats have recently been initiated. This study aimed to detect cats seropositive for Leishmania spp. using different antigens. Additional studies were performed using sera from cats with Toxoplasma gondii (n=15) to evaluate cross-reactivity. Serum samples (n=113) from cats living in the town of Ara?atuba, State of S?o Paulo, Brazil, an endemic area for human and canine visceral leishmaniasis, were tested by indirect ELISA using different antigens: crude (CAG-ELISA), fucose-mannose ligand (FML-ELISA) and K39 (rK39-ELISA). Anti-Leishmania spp. antibodies were detected in 23.0% of samples evaluated by CAG-ELISA, 13.3% by FML-ELISA and 15.9% by RK39-ELISA. Only reactive sera in all three tests were considered truly positive. No disagreement occurred among the tests (p<0.05). Serum samples seropositive for toxoplasmosis tested by CAG-ELISA were negative, but one sample (6.7%) was positive for FML-ELISA and rK39-ELISA suggesting a cross-reaction between these antigens and anti-T. gondii antibodies. These findings indicate the occurrence of feline leishmaniasis in Ara?atuba. Further studies are required to clarify the role of cats in the epidemiological cycle of leishmaniasis.     

Comparative study between atropine and hyoscine-N-butylbromide for reversal of detomidine induced bradycardia in horses

Reasons for performing study: Bradycardia may be implicated as a cause of cardiovascular instability during anaesthesia. Hypothesis: Hyoscine would induce positive chronotropism of shorter duration than atropine, without adversely impairing intestinal motility in detomidine sedated horses. Methods: Ten minutes after detomidine (0.02 mg/kg bwt, i.v.), physiological saline (control), atropine (0.02 mg/kg bwt) or hyoscine (0.2 mg/kg bwt) were randomly administered i.v. to 6 horses, allowing one week intervals between treatments. Investigators blinded to the treatments monitored cardiopulmonary data and intestinal auscultation for 90 min and 24 h after detomidine, respectively. Gastrointestinal transit was assessed for 96 h via chromium detection in dry faeces. Results: Detomidine significantly decreased heart rate (HR) and cardiac index (CI) from baseline for 30 and 60 min, respectively (control). Mean ± s.d. HR increased significantly 5 min after atropine (79 ± 5 beats/min) and hyoscine (75 ± 8 beats/min). After this time, HR was significantly higher after atropine in comparison to other treatments, while hyoscine resulted in intermediate values (lower than atropine but higher than controls). Hyoscine and atropine resulted in significantly higher CI than controls for 5 and 20 min, respectively; but this effect coincided with significant hypertension (mean arterial pressures >180 mmHg). Auscultation scores decreased from baseline in all treatments. Time to return to auscultation scores ≥12 (medians) did not differ between hyoscine (4 h) and controls (4 h) but atropine resulted in significantly longer time (10 h). Atropine induced colic in one horse. Gastrointestinal transit times did not differ between treatments. Conclusion: Hyoscine is a shorter acting positive chronotropic agent than atropine, but does not potentiate the impairment in intestinal motility induced by detomidine. Because of severe hypertension, routine use of anticholinergics combined with detomidine is not recommended. Potencial relevance: Hyoscine may represent an alternative to atropine for treating bradycardia.     

INFLUENCE OF CORN (ZEA MAYS L.) CULTIVAR DEVELOPMENT ON RESIDUE PRODUCTION

The amount and composition of crop residues added to soil in agroecosystems can influence decomposition processes and soil organic matter levels. This study aimed to evaluate residues (quantity and quality) of different corn cultivars commonly used in Brazilian cropping systems. The experiment was conducted for two seasons (2005/2006 and 2006/2007) in Rolândia, Paraná State, Brazil. Ten corn cultivars that represent five degrees of breeding development (i.e., landrace, commercial variety, double cross, triple cross, and single cross hybrids) were evaluated. At harvest, carbon (C) and nitrogen (N) of non-yield residue and grain were determined. Except for grain C, other measures (grain N concentration, residue C and N concentration, and C:N ratio) varied among cultivars. In general, the hybrids had higher residue C and lower residue N concentrations than the landraces and commercial varieties. Findings suggest that breeding selection may have altered residue production and composition, which may influence soil C dynamics.     

Evaluation of Cymbopogon flexuosus and Alpinia zerumbet essential oils as biopesticides against Callosobruchus maculatus

Journal of Plant Diseases and Protection - This study aimed to investigate the effects of the essential oils of Cymbopogon flexuosus and Alpinia zerumbet in the treatment of cowpea [Vigna...