Genotyping of potentially zoonotic Giardia duodenalis from exotic and wild animals kept in captivity in Brazil

We have studied the variability of glutamate dehydrogenase (gdh) and small subunit ribosomal (SSU) rRNA coding genes of Giardia species in fecal samples isolated from wild and exotic animals in Brazil, and compared with homologous sequences of isolates from human and domestic animals characterized in previous studies. Cysts of Giardia duodenalis were obtained from feces of naturally infected monkeys (Alouatta fusca) (n=20), chinchillas (Chinchilla lanigera) (n=3), ostriches (Struthio camelus) (n=2) and jaguar (Panthera onca) (n=1). Assemblage AI was assigned to the unique isolate of jaguar. All the samples from monkeys, chinchillas, and ostriches were assigned to Assemblage B. There was little evolutionary divergence between the referred isolates and isolates described elsewhere. The Assemblage B isolates identified in this study were closely related to Assemblage BIV isolated from humans. The molecular identification of Assemblages A and B of G. duodenalis isolates from exotic and wild animals demonstrates that such hosts may be a potential reservoir for zoonotic transmission of G. duodenalis.     

Long-term productivity growth and sustainability of Greek irrigated agriculture

The present study examines the long-term productivity growth and sustainability of Greek irrigated agriculture. A log linear trend model was used to measure long-term productivity growth, and the total factor productivity (TFP) approach measures the sustainability of the irrigated production system. A time series database of inputs and outputs data were used for the production period of 1961?–?2001 (41 years) which revealed that there is an increasing trend in the irrigated area of the country. The annual growth rate of the irrigated area is 2.6% during this time period. The long-term trends in yields of crops grown in irrigated agriculture are positive. Fertilizer use rate per ha sharply increases during this period. The productivity of irrigated Greek agriculture also increases over the time period as a result of the introduction of High Yielding Variety (HYV), seed, fertilizer and water technology. The measurement of TFP indices indicates that the long-term trend in productivity growth of irrigated Greek agricultural system is positive. The result implies that the irrigated Greek agricultural production system has long-term sustainability. The result also indicates that fertilizer, fixed capital, rainfall and temperature are positively contributing to the mean production of Greek irrigated agriculture. On the other hand, pesticide, labour and fixed capital reduces variability of irrigated agriculture. To this end, long-term agricultural sustainability depends on patterns of input use.     

Inhibition of Trypanosoma cruzi by plant extracts used in Chinese medicine

In this work, we assessed the effect of extracts obtained from 17 plants used in traditional Chinese medicine. These extracts were tested in vitro with the epimastigote form of Trypanosoma cruzi, clone Bra C(15) C(2), at 27 degrees C in F-29 medium at a concentration of 100 microg/ml in axenic cultures. Allopurinol was used as reference drug. Seven plant extracts showed inhibitory activities lower than 25%. Pueraria lobata, Mahonia beaei, Dictamus dasycarpus, Kochia scoparia, Sophora flavescens and Ligustrum lucidum showed effects with inhibition values between 25% and 60%, whereas Lithospermum erythrorhizon, Saussurea lappa, Melia toosendan and Cinnamomum cassia showed the greatest inhibitory activity of 100%. The IC(50) of these extracts were: 0.4, 2.4, 1.8 and 3.9 microg/ml, respectively. The MTT assay was made and did not show cytotoxic activity. These results allowed us to suggest that L. erythrorhizon, S. lappa, M. toosendan and C. cassia could be a source of new compounds against T. cruzi.     

Seroprevalence and isolation of Toxoplasma gondii from free-range chickens from Espírito Santo state, southeastern Brazil

Prevalence of Toxoplasma gondii infection in 510 free-range (FR) chickens (380 from 33 small farms, and 130 from a slaughter house for FR chickens) from Espírito Santo state, southeastern Brazil, was investigated. Antibodies to T. gondii were sought using commercial indirect haemagglutination (IHAT, Imuno-HAI Toxo(?), Wama Diagnóstica, S?o Paulo, Brazil, cut-off 1:16) and the modified agglutination test (MAT, cut-off 1:25) tests. Attempts were made to isolate viable T. gondii from seropositive chickens by bioassay in mice. Pooled samples of brain, heart and quadriceps muscle of one thigh (total 40 g) from 64 chickens with IHAT titers of ≥ 1:16 were minced, digested in pepsin and bioassayed in mice. Antibodies to T. gondii were found in 40.4% (206/510) FR chickens by IHAT (titer ≥ 1:16) and 38.8% (198/510) by MAT (titer ≥ 1:25); concordance between IHAT and MAT was 81.6% (kappa index=0.614). Viable T. gondii was isolated (designated TgCkBr234-281) from 48 of 64 (75%) seropositive (IHAT titers ≥ 1:32) FR chickens. Most isolates of T. gondii were virulent for mice; 100% of mice inoculated with 44 of 48 isolates died of toxoplasmosis within 30 days post inoculation (p.i). An epidemiological investigation revealed that people living in rural areas have little knowledge about the parasite and about the risk of acquiring it from raw meat. Results indicated that the locally available IHAT was useful for screening of chicken sera for T. gondii antibodies.     

Prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in sheep from Mossoró, Rio Grande do Norte, Brazil

Toxoplasma gondii is an obligate intracellular parasite with a variety of hosts, responsible for reproductive problems and economic losses in sheep flocks. Neospora caninum was recently identified and its clinical presentation in sheep is similar to that of toxoplasmosis, which can cause repeated abortions, though less frequently in this species. In order to confirm the prevalence of these agents in the city of Mossoró, Rio Grande do Norte, Brazil, 409 serum samples from adult sheep (364 females and 45 males) were tested by the indirect immunofluorescence antibody test, using cut-off point at a dilution of 1:64 and 1:50 for T. gondii and N. caninum, respectively. From the 35 properties examined, 23 (65.7%) had at least one seropositive animal for T. gondii and six (17.1%) for N. caninum. The prevalence of seropositive animals for T. gondii was 20.7% and for N. caninum 1.8%. There was no association between the presence of the agent’s antibody and gender, reports of reproductive problems and presence of dogs and/or cats in the properties. T. gondii is well distributed and N. caninum has low prevalence in sheep and in the properties of the studied region.     

Third‐compartment cannulation in alpacas using a polyurethane gastrostomy tube

Objective To develop a simple and effective surgical technique for third‐compartment cannulation in alpacas. Design Prospective study using six adult male alpacas. Methods General anaesthesia was induced and a polyurethane gastrostomy tube was surgically implanted into the distal portion of the third compartment. Results Three of the alpacas retained their cannulas for a 100‐day period; however, three cannulas were dislodged during the study. Two of the three dislodged cannulas were replaced during a second surgical procedure. Cannulas were well tolerated by the alpacas and all animals remained clinically healthy during the study period. Third compartment contents did not leak from the cannulation site. The tubes were manually removed following the completion of the study and the small defect in the body wall quickly healed over in all animals. Conclusion Surgical placement of polyurethane tubes designed for percutaneous endoscopic gastrostomy is a useful method of cannulating the third compartment in camelids. This technique can be used for experimental studies and possibly could be used for nutritional support and fluid therapy in sick camelids that might need long‐term care.     

Molecular mechanism of Chinese medicine formula (Bushen Bugu Tang) for the treatment of postmenopausal osteoporosis

AIM: To study the molecular mechanism of Chinese medicine formula Bushen Bugu Tang (BSBGT) for the treatment of postmenopausal osteoporosis (PMO). METHODS: Forty female SD rats were divided into four groups: the sham group, OVX group, estrogen treatment group and BSBGT treatment group, ten for each, difference operation and treatment were given. Subsequently, bone mineral density (BMD), bone histomorphometric changes and collagen type I expression in the femur of these rats were examined. In order to position the target of the formula acting on bone, the effect of BSBGT on regulation of mRNA levels of RANKL and OPG, two key factors for regulating osteoclastogenesis, by using rat calvarial-derived stromal cell cultrues, was elucidated. RESULTS: The BSBGT formula was found as effective as estrogen in preventing the reduction of BMD, the trabecular area and trabecular numbers in femora and also the decrease in collagen type I expression following ovariectomy. RT-PCR resulted showed that both estrogen and BSBGT significantly up-regulated OPG mRNA level and down-regulated RANKL mRNA level, resulting in an increased ratio of OPG/RANKL in cultured rat stromal cells. CONCLUSIONS: The ratio of OPG/RANKL within the osseous environment determines whether bone formation predominates bone resorption. We therefore concluded that the BSBGT formula might be as effective as estrogen in prevention of postmenopausal osteoporosis through the mechanism of up-regulating RANKL expression and down-regulating OPG expression in osteoblast/stromal cells.     

Occurrence of Neospora caninum antibodies in sera from dogs of the city of São Paulo,Brazil

Neospora caninum is an important cause of abortion in dairy cattle worldwide. Dogs are important in the epidemiology of this parasite because they are the only hosts known to excrete N. caninum oocysts. In order to understand the prevalence of N. caninum in dogs, sera from 500 owned dogs and from over 600 feral street dogs from the city of S?o Paulo, Brazil were assayed for antibodies to N. caninum. Sera were examined by the Neospora agglutination test (NAT) using mouse-derived tachyzoites. Antibodies (> or =1:25) to N. caninum were found in nearly 10% (49/500) of owned dogs and in 25% (151/611) of stray dogs. NAT titers for owned dogs were 1:25 in 28 (5.6%) dogs, 1:50 in 20 (4%) dogs, and > or =1:500 in 1 (0.2%) dog. NAT titers for stray dogs were 1:25 in 79 (12.9%) dogs, 1:50 in 68 (11.1%) dogs, and > or =1:500 in 4 (0.6%) dogs. These data indicate that feral dogs may be important in the epidemiology of N. caninum infection.