Biological activity of cinnamaldehyde, citronellal, geraniol and anacardic acid on Haemonchus contortus isolates susceptible and resistant to synthetic anthelmintics.

Parasitism by gastrointestinal nematodes is a challenge for small ruminant farming worldwide. It causes productive and economic losses, especially due to parasite resistance to conventional anthelmintics. Natural compounds with antiparasitic activity are a potential alternative for controlling these parasites especially when considering the widespread occurrence of anthelmintic resistance. Our objective was to evaluate the activity of anacardic acid, geraniol, cinnamaldehyde and citronellal on Haemonchus contortus isolates with different levels of anthelmintic resistance profiles. These compounds were tested using egg hatch assays (EHAs), larval development tests (LDTs) as well as LDTs on mini-fecal cultures, on the Haemonchus contortus isolates Kokstad (KOK-resistant to all anthelmintics), Inbred-Strain-Edinburgh (ISE-susceptible to all anthelmintics) and Echevarria (ECH-susceptible to all anthelmintics). Effective concentrations to inhibit 50% (EC50) and 95% (EC95) of egg hatching and larval development were calculated. Results for EHA and LDT for all tested compounds, considering EC50 and EC95 values, showed low variation among the studied isolates with most RF values below 2x. All studied compounds showed efficacy against egg hatching and larval development of H. contortus isolates regardless of anthelmintic resistance profiles. The compounds with the smallest EC50 and EC95 values were cinnamaldehyde and anacardic acid making them promising candidates for future in vivo studies.

Castor bean cake for the control of parasites in pasture-finished sheep.

The purpose of this study was to evaluate castor bean cake as alternative input in the control of gastrointestinal parasites in sheep raised on irrigated pasture under continuous stocking. The treatments consisted of sheep supplemented with standard diet and pasture fertilized with urea; sheep supplemented with alternative diet and pasture fertilized with urea; sheep supplemented with standard diet and pasture fertilized with in natura castor bean cake; and sheep supplemented with alternative diet and pasture fertilized with in natura castor bean cake. A randomized complete block design (CBD) was used, with 16 replications (sheep), with repeated measures over time, the plots being the treatments, and the subplots the collection times. Infective nematode larvae in the pasture (L3.g DM-1), number of eggs per gram of feces (EPG), globular volume (GV), and total plasma protein (TPP) were evaluated. For adult gastrointestinal parasite counts, CBD was also used with six replications. Except for FAMACHA© grade, all variables had effect (P < 0.01) of the time factor. The average number of L3.g DM-1 and EPG were 126 and 841, respectively, with no effect (P > 0.05) of the treatment factor. The values observed for GV and TPP were higher than 25.9% and 6.4 g·dL-1, respectively, which were considered normal. As organic fertilizer, the fractionated application of in natura castor bean cake does not reduce the contamination of pastures by nematode larvae. The evaluated feeds improve the resilience of the sheep to infection by gastrointestinal parasites. The treatments using castor bean cake reduced the adult parasites present in the abomasum of sheep.

Quantitative molecular diagnosis of levamisole resistance in populations of Haemonchus contortus.

Parasitism by Haemonchus contortus is one of the main limiting factors in small ruminant production around the globe. Although several studies suggest the use of integrated management practices, these parasites have been controlled essentially with synthetic anthelmintic drugs. The resistance mechanism against the imidazothiazole derivative levamisole in Haemonchus contortus has not been fully described. Recently, resistance was associated with a 63bp deletion in the Hco-acr-8b gene that encodes a subunit for a nicotinic acetylcholine receptor. This study aimed to standardize a real time PCR (qPCR) protocol for levamisole resistance diagnosis in H. contortus populations based on this polymorphism and use it to characterize 23 field H. contortus populations obtained from different localities of Ceará State, Northeast Brazil. In addition, two populations of H. contortus were used as a standard of susceptibility and resistance, Inbred Strain Edinburgh (ISE) and Kokstad, respectively. Larval development tests (LDT) were performed on five field isolates and both EC50 and EC95 were estimated. LDT EC95 values provided a wider interval between susceptible and resistant populations than EC50 values (EC95 = 1.96-57.93 µM; EC50 = 0.05-0.39 µM), and were found to be more appropriate for differentiating them. Real time PCR results showed resistance allele frequencies ranged from 20.9 to 76.7%. Our results suggest that levamisole resistance may be present in field populations but it is not as widespread as benzimidazole resistance. This methodology may be useful to monitor levamisole resistance in field populations of H. contortus.

Strategic vaccination of hair sheep against Haemonchus contortus.

In this study, we evaluated in two trials a protocol designed to protect hair sheep using Barbervax®, a vaccine containing Haemonchus contortus gut membrane glycoprotein antigens. Results indicated that naturally infected vaccinated sheep had significant egg count reductions (90.2 ± 4.03%) compared with controls, although blood parameters remained relatively unchanged probably because the level of challenge was low. Vaccination prevented the periparturient rise in egg shedding of ewes, as well as egg shedding in lambs (37.1%). In the second trial, sheep which were experimentally exposed to higher artificial challenge also showed an efficient response to the vaccine as confirmed by high antibody levels and reduced egg counts and worm burdens (87 ± 5.4% and 79%) respectively. Thus, we believe that the vaccine should be integrated with other management practices for meat hair sheep as it has the advantages of adequate efficacy, reducing anthelmintic utilization and avoiding milk and environmental contamination with chemical residues.

Infectious diseases during pregnancy in Brazil: seroprevalence and risk factors.

INTRODUCTION: Vertically transmitted infections are caused by a diversity of pathogenic microorganisms. Pregnant women are routinely screened to evaluate the risks and reduce the burden of disorders in their unborn children. We assessed the prevalence and possible risk factors for Cytomegalovirus (CMV), Rubella, Human T lymphotropic virus (HTLV), and Toxoplasma gondii in pregnant women from the South region of Bahia State, Brazil. METHODOLOGY: Serum samples were obtained from 726 pregnant women aged between 13 and 44 years, with a median age of 24 years. ELISA assays were used to detect CMV, Rubella, HTLV and T. gondii IgG and IgM antibodies. RESULTS: The prevalence rates of IgG antibodies found were 95.2% for CMV, 97.0% for Rubella, and 72.3% for T. gondii. Furthermore, the prevalence of HTLV-1/2 was 1.2%. IgM antibodies were reactive only for CMV (0.8%) and T. gondii (3.7%). Variables independently associated with the detection of anti-T. gondii IgG antibodies were white self-reported race/ethnicity (Odds Ratio [OR] 2.26, 95% CI 1.26-4.06, P = 0.006), wage income (OR 0.55, 95% CI 0.35-0.88, P = 0.013), and history of previous pregnancy (OR 1.60, 95% CI 1.02-2.50, P = 0.038). CONCLUSIONS: This study highlights the importance of monitoring for infectious diseases during pregnancy and initiation of early interventions to reduce the burden of fetal losses and other important infant sequelae attributable to congenital infections.

Haemonchus contortus ß-tubulin isotype 1 gene F200Y and F167Y SNPs are both selected by ivermectin and oxfendazole treatments with differing impacts on anthelmintic resistance.

Parasitism by Haemonchus contortus is one of the main limiting factors in small ruminant production in tropical areas. Benzimidazoles (BZ) and macrocyclic lactones (ML) are the most used anthelmintic classes in gastrointestinal nematodes control. There is considerable scientific evidence of a possible relation between the anthelmintic resistance to BZ and ML. This study aimed to characterize the dynamics of anthelmintic resistance in an H. contortus susceptible isolate under selection pressure for BZ and ML alone or in combination and the role of isotype 1 ß-tubulin gene SNPs in these situations. A total of 12 Somali sheep were infected with 5000 third stage larvae of H. contortus Inbred-Susceptible Edinburgh (ISE) isolate. Once infection was established, animals were distributed in three groups (n=4), each treated with crescent doses of oxfendazole (OXF), ivermectin (IVM) and oxfendazole plus ivermectin (IVMOXF). An additional control group with untreated animals was maintained during the entire experiment. After each treatment, eggs were collected and real-time PCR was performed to identify single nucleotide polymorphisms (SNPs) F167Y, F200Y and E198A, in addition to egg hatch test (EHT) for BZ and larval development test (LDT) for ivermectin resistance. All treatments led to increased resistance allelic frequencies at SNPs F200Y and F167Y (p <0.05). In vitro results showed increased phenotypic resistance against both anthelmintic classes in groups IVM and IVMOXF while group OXF only developed resistance against BZ. Finally, we provide evidence that while isotype 1 ß-tubulin gene SNPs may have some involvement with ML resistance, the presence of these ß-tubulin SNPs alone are not sufficient to develop ML resistance.

Coccidia of gallinaceous meat birds in Brazil.

Coccidiosis is a disease that limits the production and marketing of gallinaceous birds in North America, especially quails, pheasants and chukar partridges. Virtually no research has been conducted in South America on the causative agents of diseases among these birds, including coccidia. The aim of this work was to make first observations on Eimeria spp. in the chukar partridge Alectoris chukar and the grey quail Coturnix coturnix, which are reared for meat in Brazil. Fecal and tissue samples were collected from commercial farms and were examined for oocysts, gross and microscopic lesions or endogenous stages. From this examination, it was found that partridges raised in Brazil did not have any visible infection. However, grey quails presented mild infection and two Eimeria species that had previously been described in other birds were identified.

Coccidia of gallinaceous meat birds in Brazil / Coccídios de galináceos de corte no Brasil

Coccidiosis is a disease that limits the production and marketing of gallinaceous birds in North America, especially quails, pheasants and chukar partridges. Virtually no research has been conducted in South America on the causative agents of diseases among these birds, including coccidia. The aim of this work was to make first observations on Eimeria spp. in the chukar partridge Alectoris chukar and the grey quail Coturnix coturnix, which are reared for meat in Brazil. Fecal and tissue samples were collected from commercial farms and were examined for oocysts, gross and microscopic lesions or endogenous stages. From this examination, it was found that partridges raised in Brazil did not have any visible infection. However, grey quails presented mild infection and two Eimeria species that had previously been described in other birds were identified.

A coccidiose é uma enfermidade limitante para a produção e comercialização de aves de corte na América do Norte, principalmente codornas, faisões e a perdiz de chukar. Praticamente nenhuma pesquisa foi realizada na América do Sul sobre os agentes causadores de doenças nessas aves, incluindo coccídios. O objetivo deste trabalho foi realizar as primeiras observações sobre Eimeria spp. em perdiz de chukar Alectoris chukar e codornas cinzentas Coturnix coturnix criadas para abate no Brasil. Amostras de fezes e de tecidos foram coletadas em granjas comerciais e examinadas para oocistos, lesões macroscópicas e microscópicas ou estágios endógenos. Após o exame, verificou-se que a criação de perdizes, no Brasil, não tinha infecção visível. No entanto, as codornas cinzentas apresentaram uma infecção leve e foram identificadas duas espécies de Eimeria descritas anteriormente em outras aves.

Ascaridiasis in peafowl Pavo cristatus (Phasianidae) due to Ascaridia galli Schrank, 1788.

Twelve white peafowl (Pavo cristatus) affected by an outbreak of an intestinal disease were referred for more detailed examination at the Universidade Estadual de Santa Cruz-BA, Brazil. During the course of the disease, peachicks were severely affected, with enteric signs such as diarrhea plus dehydration, decreased feed intake and progressive weight loss. After examination, 8 of 12 samples (66.6%) presented single or mixed nematode infection and Ascarid eggs were the most frequent finding on fecal examination. Adult peafowl did not present clinical signs even when positive after fecal exam. Morphological analysis, clinical signs, fecal and gross examinations resulted in a diagnosis of ascaridiasis caused by Ascaridia galli Schrank (1788).

Eimeria species in dairy goats in Brazil.

The focus of this work is to determine the distribution and identify species of Eimeria parasites of dairy goats in the livestock of the National Goat and Sheep Research Center in Sobral, State of Ceará, Northeast Brazil. Results showed the presence of multiple species in 196 of 215 analyzed samples (91.2%). Fifty five out of these were from kids (28%) and 141 from adult goats (72%). Eight different Eimeria species were identified and their prevalence in the herd was: Eimeria alijevi Musaev, 1970 (26.7%), E. arloingi (Marotel, 1905) Martin, 1909 (20.6%), E. hirci Chevalier, 1966 (18%), E. ninakohlyakimovae Yakimoff & Rastegaieff, 1930 (16.2%), E. jolchijevi Musaev, 1970 (8.7%), E. christenseni Levine, Ivens & Fritz, 1962 (6%), E. caprovina Lima, 1980 (2.8%) and E. caprina Lima, 1979 (1%). Moreover, E. ninakohlyakimovae showed higher prevalence in kids (97%), followed by E. arloingi and E. alijevi (88%). On the other hand, E. alijevi (77%) was more common in adult goats followed by E. hirci (74%) and E. ninakohlyakimovae (70%). The species E. caprina had low frequency in both kids (27%) and adult goats (13%). Data indicated that infection was relatively common among kids and adult goats. The implementation of a routine diagnostic strategy can be useful in maintaining Eimeria populations under monitoring and will enable the determination of its potential impact on dairy goat herds in Northeast Brazil.

Diagnosis of Eimeria species using traditional and molecular methods in field studies.

The objective of this study was to identify and characterize species of Eimeria in broiler chickens using traditional morphological and pathological plus molecular (DNA amplification) diagnostic methodologies. Using a combination of those techniques it was possible to identify the presence of multiple circulating species in the flock as well as higher frequencies for some of them, especially Eimeria praecox and Eimeria maxima, which were identified in 100% of the flocks. The frequencies of the other species were Eimeria mitis and Eimeria necatrix (93.3%), Eimeria tenella (76,7%), Eimeria acervulina (56.7%) and Eimeria brunetti (16.7%). However using the lesion score, the most common species were E. maxima (46.7%), E. acervulina (30%), E. tenella (23.3%), and E. necatrix (10%). E. brunetti and E. praecox were not identified by using lesion score. DNA amplification had detection sensitivity for Eimeria species in the field samples of at least 20 oocysts. The implementation of DNA amplification as a routine diagnostic technique in aviaries can assist Eimeria population.

Tyzzeria parvula (Kotlan, 1933) Klimes, 1963 (Apicomplexa: Eimeriidae) in the greylag goose (Anser anser Linnaeus, 1758) in southeastern Brazil.

Tyzzeria parvula from the greylag goose (Anser anser) is described in Southeastern Brazil. Oocysts are spherical to subspherical ranging from 12.4-180 to 10.7-15.9 microm (15.4 X 13.4 microm), shape index 1.15, with a double layered wall 0.4 to 0.7 microm thick (n=20), outer smooth and colorless, inner pale greenish. Micropyle is absent, but oocyst residuum is present containing numerous granules and spherules. Surrounded by residuum there are eight sporozoites having one end round and other fine and slightly curved.

Alterações clínicas e patológicas durantea infecção experimental com Eimeria acervulinaem frangos de corte suplementados com betaína / Clinical and pathological changes during the experimental infection with Eimeria acervulina in broiler chickens suplemented with betaine

Com o objetivo de se avaliar o efeito da betaína na patogenicidade de Eimeria acervulina, 420 pintos de corte Cobb foraminoculados experimentalmente com 2 x 105 oocistos esporulados e alocados em baterias num esquema de blocos ao acasocom cinco tratamentos e seis repetições, incluindo-se um controle positivo, um grupo tratado com o anticoccidiano salinomicinae promotor de crescimento, e mais três níveis de betaína como aditivo na ração a 0,05, 0,10 e 0,15 por cento. No 14o dia de vida as avesforam individualmente infectadas com 2 x 105 oocistos esporulados. A betaína não foi capaz de interferir positivamente naresposta clínica e patologia das aves durante a infecção por E. acervulina, no entanto foi capaz de aumentar a produção deoocistos. É provável que a betaína possa ser utilizada como aditivo na ração das aves em substituição aos medicamentosconvencionais salinomicina e penicilina G potássica, cujos efeitos são semelhantes na recuperação do hospedeiro frente àcoccidiose.

Purposing to investigate the betaine effect on biology and morphology of developmental stages of Eimeria acervulina, 420 Cobbbroiler chicks were experimentally inoculated with 2 x 105 sporulated oocysts and housed in battery cages in a block design withfive treatments and six replicates each, including a positive control, a group treated with the anticoccidial salinomycin plusantibiotic growth promoter plus three levels of betaine as additive in the feed 0.05, 0.10 and 0.15 percent. Betaine was not able to affectthe clinical response and pathology during the infection, but improved oocyst output, possibly due to the osmotic mechanisms.In fact betaine could replace salinomycin plus growth promoter as additive in the feed which effects is closer in the recovery ofthe host affected by coccidiosis.

Alterações clínicas e patológicas durante a infecção experimental com Eimeria acervulina em frangos de corte suplementados com betaína

Com o objetivo de se avaliar o efeito da betaína na patogenicidade de Eimeria acervulina, 420 pintos de corte Cobb foraminoculados experimentalmente com 2 x 105 oocistos esporulados e alocados em baterias num esquema de blocos ao acasocom cinco tratamentos e seis repetições, incluindo-se um controle positivo, um grupo tratado com o anticoccidiano salinomicinae promotor de crescimento, e mais três níveis de betaína como aditivo na ração a 0,05, 0,10 e 0,15%. No 14o dia de vida as avesforam individualmente infectadas com 2 x 105 oocistos esporulados. A betaína não foi capaz de interferir positivamente naresposta clínica e patologia das aves durante a infecção por E. acervulina, no entanto foi capaz de aumentar a produção deoocistos. É provável que a betaína possa ser utilizada como aditivo na ração das aves em substituição aos medicamentosconvencionais salinomicina e penicilina G potássica, cujos efeitos são semelhantes na recuperação do hospedeiro frente àcoccidiose.

Eimeria auritanensis n. sp. and E. gambai Carini, 1938 (Apicomplexa: Eimeriidae) from the opossum Didelphis aurita Wied-Newied, 1826 (Marsupialia: Didelphidae) from southeastern Brazil.

Two eimerid species are described from the opossum Didelphis aurita from southeastern Brazil. Eimeria auritanensis n. sp. sporulated oocysts spherical to subspherical (31.55 +/- 1.56 by 29.55 +/- 1.40 microm), shape index 1.07; oocyst wall double layered 2.10 +/- 0.27 microm thick, outer yellowish and strongly ornamented having a prominently mammillated surphace; inner layer smooth and brownish. There is no micropyle or oocyst residuum, but one or two polar granules are present. Sporocysts ovoid (13.20 +/- 1.64 by 10.41 +/- 1.10microm) with a faint Stieda's body and residuum composed of granules and spherules. Eimeria gambai Carini, 1938 sporulated oocysts ovoid to subspherical (26.54 +/- 1.7 by 24.82 +/- 1.85 microm), shape index 1.07; oocyst wall double layered 2.10 +/- 0.27 microm thick, outer colorless to pale yellow entirely pitted, while inner layer smooth dark yellow to pale brow. Micropyle and residual bodies absent, polar granules present. Sporocysts ovoid 12.49 +/- 1.75 by 9.32 +/- 1.01 microm. Stieda body present and round. Sporocysts residuum composed of many granules and spherules.

[The effect of betain on biology and morphology of developmental stages of Eimeria acervulina in broiler chicks experimentally infected]. / Efeito do uso da betaína na biologia e morfologia dos estádios evolutivos de Eimeria acervulina em frangos de corte infectados experimentalmente com oocistos esporulados.

Purposing to investigate the betaine effect on biology and morphology of developmental stages of Eimeria acervulina, 420 broiler chicks Cobb were experimentally inoculated with 2 x 10(5) sporulated oocysts and housed in battery cages in a block design with five treatments and six replicates each, including a positive control, a group treated with salinomycin and growth promoter plus three levels of betaine as additive in the feed at 0.05, 0.10 and 0.15%. Measurements of oocysts, sporocysts and endogenous stages were performed as morphological parameters, while pre patent and patent periods and sporulation time were taken as biological parameters. Morphology was also associated with the mathematical constant Phi (1.618) to evaluate possible relationship. Betaine was able to cause modifications in both biology and morphology of oocysts and sporocysts, whereas it was weakly able to affect developmental stages based on trophozoites and macrogamonts measurements. According to the measures of sporocysts E. acervulina development was closely related to Phi.

Species of the genus Eimeria (apicomplexa: Eimeriidae) from japanese quails (Coturnix japonica) in Brazil and E. fluminensis for the preoccupied E. minima of this quail

Amostras de fezes de 120 codornas japonesas de uma criaçäo localizada no estado do Rio de Janeiro, Brasil, foram submetidas à técnica de centrífugo-flotaçäo em soluçäo saturada de açúcar para identificar quantas espécies do gênero Eimeria possam ser encontradas parasitando codornas. Quatro espécies distintas foram identificadas como Eimeria bateri (Bhatia, Pandey e Pande, 1965), onde os oocistos mediam 22,28ñ2,70 por 18,38ñ2,50µm; E. uzura Tsunoda and Muraki, 1971 com 20,82ñ1,78 por 16,70ñ1,34µm; E. tsunodai Tsutsumi, 1972 com 19,85ñ1,93 por 15,55ñ1,50µm, e E. fluminensis como espécie para a espécie pré-ocupada E. minima Teixeira, Lopes, 2000 da codorna japonesa com 17,31ñ1,87 por 16,45ñ1,72µm de diâmetros, respectivamente. A descriçäo de cada espécie foi feita com base nos aspectos morfológicos dos oocistos, após esporulaçäo em uma soluçäo de 2,5 por cento de bicromato de potássio a 27ºC.

Eimeria minima n. dp. (Apicomplexa: Eimeriidae) from the Japanese quail (Cuturnix cuturnix japonica) in Brazil

Oocysts from feces of Japanese quails (Cuturnix cuturnix japonica) were described. Sporulated oocysts were spherical to subspherical and measured 17.31ñ1.87 to 16.45ñ1.72µm in size with shape index 1.05ñ0.04. The oocyst wall was thin and smooth, and was consisted of two distinct layer, outside layer was brownish while the inner one was hyaline. Few polar granules were present but without a micropyle or an oocyst residuum. Sporocysts were ovoid with 10.43ñ1.11 to 5.92ñ0.58 µm in diameter. Sporocyst residuum was present with visible granules with tactful Stieda body. Each sporocyst had two sporozoites. For the sake of small size in comparison to the others species found in the Japanese quail, Eimeria minima was proposed.