Benefits and costs of immune memory in Rhodnius prolixus against Trypanosoma cruzi.

There is increasing evidence supporting the immune memory in invertebrates, but the studies are relatively neglected in insect vectors other than mosquitoes. Therefore, we tested two hypotheses: 1) Rhodnius prolixus insects possess immune memory against Trypanosoma cruzi, and 2) their immune memory is costly. The Dm28c and Y strains of T. cruzi were used, the former being more infective than the latter. On the one hand, the triatomines subjected to dual challenges with the Dm28c strain did not show significant differences in survival than those of the heterologous challenge groups control-Dm28c and Y-Dm28c. On the other hand, the insects survived longer after a dual Y-Y challenge than after the corresponding heterologous challenge (control-Y). The Y-Y, Dm28c-Y, and naïve groups showed similar survival. There was more prolonged survival following the Y-Y versus Dm28c-Dm28c dual challenge. The Dm28c-Dm28c group exhibited moulting sooner than the control-Dm28c or naïve group. In contrast, there were no differences in the probability of moulting between the Y-Y and naïve groups. The results suggest that triatomines have immune memory against the Y but not the Dm28c strain. Further investigation on triatomine and T. cruzi interaction is needed to determine if infectivity accelerates or delay growth due to innate immune memory.

Genetic diversity of Bm86 sequences in Rhipicephalus (Boophilus) microplus ticks from Mexico: analysis of haplotype distribution patterns.

BACKGROUND: Ticks are a problem for cattle production mainly in tropical and subtropical regions, because they generate great economic losses. Acaricides and vaccines have been used to try to keep tick populations under control. This has been proven difficult given the resistance to acaricides and vaccines observed in ticks. Resistance to protein rBm86-based vaccines has been associated with the genetic diversity of Bm86 among the ectoparasite's populations. So far, neither genetic diversity, nor spatial distribution of circulating Bm86 haplotypes, have been studied within the Mexican territory. Here, we explored the genetic diversity of 125 Bm86 cDNA gene sequences from R. microplus from 10 endemic areas of Mexico by analyzing haplotype distribution patterns to help in understanding the population genetic structure of Mexican ticks. RESULTS: Our results showed an average nucleotide identity among the Mexican isolates of 98.3%, ranging from 91.1 to 100%. Divergence between the Mexican and Yeerongpilly (the Bm86 reference vaccine antigen) sequences ranged from 3.1 to 7.4%. Based on the geographic distribution of Bm86 haplotypes in Mexico, our results suggest gene flow occurrence within different regions of the Mexican territory, and even the USA. CONCLUSIONS: The polymorphism of Bm86 found in the populations included in this study, could account for the poor efficacy of the current Bm86 antigen based commercial vaccine in many regions of Mexico. Our data may contribute towards designing new, highly-specific, Bm86 antigen vaccine candidates against R. microplus circulating in Mexico.

The innate immune response of triatomines against Trypanosoma cruzi and Trypanosoma rangeli with an unresolved question: Do triatomines have immune memory?

The present work aimed to review the immune response from different triatomines against Trypanosoma cruzi and Trypanosoma rangeli and propose the study of immune memory in such insects. Trypanosoma use triatomines as vectors to reach and infect mammals. A key question to be answered about vector-parasite interaction is why the immune defense and resistance of the insect against the parasites vary. Up to date data shows that the defense of triatomines against parasites includes cellular (phagocytosis, nodulation and encapsulation) and humoral (antimicrobial peptides, phenoloxidase and reactive oxygen and nitrogen species) responses. The immune response varies depending on the triatomine species, the trypanosome strain and species, and the insect intestinal microbiota. Despite significant advances to understand parasite-insect interaction, it is still unknown if triatomines have immune memory against parasites and if this memory may derive from tolerance to parasites attack. Therefore, a closer study of such interaction could contribute and establish new proposals to control the parasite at the vector level to reduce parasite transmission to mammals, including men. For instance, if immune memory exists in the triatomines, it would be interesting to induce weak infections in insects to find out if subsequent infections are less intense and if the insects succeed in eliminating the parasites.

Rat embryo quality and production efficiency are dependent on gonadotrophin dose in superovulatory treatments.

The present study was performed to optimize a superovulation protocol in rats in order to produce a large number of good-quality embryos suitable to develop rat embryonic stem (rES) cells. We first evaluated the ovulation kinetics of three rat strains: Wistar, Fisher and ACI/N. Animals (n=30 per strain) were treated with 50 IU of pregnant mare serum gonadotrophin (PMSG), and ovulation was induced with 50 IU of human chorionic gonadotrophin (hCG) 50 h apart. Next, we evaluated the dose-response curves of PMSG and hCG in Wistar rats in order to obtain the highest number of embryos. The parameters evaluated for superovulation efficiency were: percentage of mated females, percentage of pregnant females and the average number of embryos collected per female. The results of these experiments suggested that the best dose combination was 50 IU for each hormone. Subsequent experiments, again with Wistar rats, were designed to test which of four hormonal combination treatments (30/30, 30/50, 50/30, and 50/50 IU of PMSG/hCG) will produce the largest numbers of good-quality embryos. Embryo quality was evaluated by embryo development uniformity, embryo morphology, embryo survival in an in vitro culture and embryo ability to generate rES-like cells. Results from these experiments showed that 30/50 IU of PMSG/hCG was the treatment that induced the best embryo quality. In conclusion, our results indicated that, in Wistar rats, the most appropriate hormonal combination dose for superovulation protocols with high number of good-quality embryos was 30 IU of PMSG and 50 IU of hCG given 50 h apart. We are performing further studies with rES-like cells produced with the present methodology to evaluate if they are able to participate in the production of germ-line chimeras.

Sheep management and cryptosporidiosis in central Mexico.

To establish the association between sheep farm management practices and cryptosporidiosis in Central Mexico, 37 farms were invited and questionnaires answered by owners. Faeces samples were taken from lambs under 3 months old and ewes in the farms. Parasites were identified by modified Ziehl-Neelsen stain. Questionnaire included: general farm information, facilities characteristics, facilities cleaning measures, water use and farm and animal management practices. Farms were grouped in a low ( Pound 3,37%), medium (3.38-49.9%) and high basis ((3)50%) according to prevalence. Associations (OR) were calculated according to positivity, compared by chi-square test. A general prevalence of 34,33% was found. Five variables showed statistical significance (p<0.05). Associations found were: use of bed, grazing place, and bottle cleaning. The only association found between management practices and cryptosporidiosis that could represent a risk was the grazing place. This report is the first one in the country regarding predisposing conditions for cryptosporidiosis.

Prevalence of Cryptosporidium spp. in asymptomatic sheep in family flocks from Mexico State.

To determine Cryptosporidium spp. prevalence in faeces from asymptomatic sheep, 37 family flocks in Mexico State were sampled. Data were collected by the interview method and Ziehl-Neelsen stain of faecal samples for Cryptosporidium identification. The independent group proportion hypothesis test was used for differences in prevalences between sheep farms. A 34.3% general prevalence was found, and prevalence in herds with >100 animals was significantly higher (40.6%). Correlation was found between flock size and Cryptosporidium spp. presence (odds ratio, OR, 2.57; confidence interval, IC(95%), 1.84-3.60).