Seroprevalence and associated risk factors of bovine neosporosis in the Khomas region of Namibia.

Neospora caninum is a coccidian parasite that occurs worldwide and is one of the most important causes of abortion, especially in cattle. However, no studies have been performed in Namibia to determine the N. caninum status in livestock. Therefore, this study aimed to determine the seroprevalence of N. caninum in cattle and the associated risk factors in the Khomas region of Namibia. A total of 736 sera were collected from cows in 32 farming establishments. These comprised 698 beef and 38 dairy cattle sera and were tested using a commercial enzyme-linked immunosorbent assay (ELISA) kit. Questionnaires were concurrently administered to determine possible risk factors associated with N. caninum seropositivity. A total of 42 sera were positive (all beef), giving an animal-level seroprevalence rate of 5.7%. Eight of the 32 establishments had at least one positive animal, giving a herd-level seroprevalence of 25%. There was no significant association between seropositivity and the presence of dogs, jackals, history of abortions, farm size, number of cattle or average annual rainfall. The establishments with moderate to high numbers of Feliformia were 9.8 times more likely to be seropositive to N. caninum than those with none to low levels of the former (p = 0.0245). The authors concluded that the seroprevalence level of N. caninum in the Khomas region was relatively low compared with other parts of the world and that the role of Feliformia in the epidemiology of bovine neosporosis needed to be further investigated.Contribution: Serological evidence of bovine neosporosis and the associated risk factors are reported in Namibia for the first time. This study contributes to the scientific body of knowledge on N. caninum in Africa, which is currently limited.

Changes in serum biomarkers of inflammation in bovine besnoitiosis.

BACKGROUND: Acute and chronic besnoitiosis in extensive natural-service herds can have relevant effects in the health of bulls and negative consequences in their productive performance. Recent progress has been made in order to elucidate the pathogenesis of this disease. In this context, the study of biomarkers of inflammation in serum would contribute to gaining knowledge about the physiopathology of bovine besnoitiosis. Serological biomarkers could help in early diagnosis and prognosis, as seropositive bulls may have mild or severe testicular lesions. METHODS: Herein, we have investigated the diagnostic and/or prognostic value of a panel of serum (serological) biomarkers related to inflammation, including total protein, globulin and albumin, haptoglobin (Hp), adenosine deaminase (ADA) paraoxonase-1 (PON-1) and acetylcholinesterase (AChE) in naturally and experimentally B. besnoiti-infected males classified according to different clinical phases of the disease (acute, chronic and subclinical besnoitiosis). RESULTS: Results showed a similar response pattern in these biomarkers for naturally and experimentally infected cattle, with a few relevant variations. Most significant changes occurred during the acute phase of infection, although significant changes in a few biomarkers were also observed during the chronic infection. Haptoglobin, albumin, PON-1 and ADA were identified as the biomarkers that showed changes of higher magnitude in the acute phase of the infection, whereas high total protein and globulin values were found in chronically infected cattle. We have described the changes of a panel of inflammatory biomarkers of acute and chronic bovine besnoitiosis. CONCLUSIONS: In summary, several biomarkers with promising diagnostic value have been identified. The biomarkers associated with acute infection are related to previously reported molecular biomarkers in testicular parenchyma of infected bulls and could help in the diagnosis of early infections and complement results from specific immunoglobulin M (IgM) detection.

Phenological and intrinsic predictors of mite and haemacoccidian infection dynamics in a Mediterranean community of lizards.

Ectotherms are vulnerable to environmental changes and their parasites are biological health indicators. Thus, parasite load in ectotherms is expected to show a marked phenology. This study investigates temporal host­parasite dynamics in a lizard community in Eastern Spain during an entire annual activity period. The hosts investigated were Acanthodactylus erythrurus, Psammodromus algirus and Psammodromus edwardsianus, three lizard species coexisting in a mixed habitat of forests and dunes, providing a range of body sizes, ecological requirements and life history traits. Habitat and climate were considered as potential environmental predictors of parasite abundance, while size, body condition and sex as intrinsic predictors. Linear models based on robust estimates were fitted to analyse parasite abundance and prevalence. Ectoparasitic mites and blood parasites from two haemococcidian genera were found: Lankesterella spp. and Schellackia spp. Habitat type was the only predictor explaining the abundance of all parasites, being mostly higher in the forest than in the dunes. The results suggest that particularities in each host­parasite relationship should be accounted even when parasites infect close-related hosts under the same environmental pressures. They also support that lizard parasites can be biomarkers of environmental perturbation, but the relationships need to be carefully interpreted for each host­parasite assemblage.

Multilocus Genotyping of Sympatric Hepatozoon Species Infecting the Blood of Ontario Ranid Frogs Reinforces Species Differentiation and Identifies an Unnamed Hepatozoon Species.

Intraerythrocytic gamonts of at least 2 named Hepatozoon species have been reported to infect the erythrocytes of ranid frogs in Ontario, Canada. Although gamonts of both species are morphometrically similar, the cytopathological changes that 1 of these species, Hepatozoon clamatae, causes to host erythrocytes, manifested by nuclear fragmentation, was used historically to distinguish this parasite from Hepatozoon catesbianae. Molecular characterization of these 2 Hepatozoon species has been equivocal in correlating genotype with gamont morphotype. Amplification and sequencing of multiple potential genotyping loci within the nuclear (18S ribosomal deoxyribonucleic acid [rDNA]; internal transcribed spacer 1), apicoplast (23S rDNA), and mitochondrial genomes (complete genomes, cytochrome c oxidase subunits I and III [COI and COIII], and cytochrome b) were conducted on Hepatozoon species that infect ranid frogs in Ontario. Sequence data were then used to evaluate the diversity of parasites present in these amphibian hosts and to assign genotypes to gamont morphotypes, if possible. Three distinct genotypes were identified at all loci; the data permitted the discovery of a third, formerly unrecognized Hepatozoon species in ranid frogs from Ontario. Although all genetic loci demonstrated differences between Hepatozoon species, mitochondrial COIII sequences were most suitable for genotypic differentiation of these parasites of frogs. Linking genotypes to gamont morphotypes proved impossible; genotypes identified as H. catesbianae and H. clamatae were found in infections with or without nuclear fragmentation of their host erythrocytes. This suggests that differentiating these species must rely on suitable genotyping methods for identification in the blood of their amphibian intermediate hosts.

Exploring alterations in hematological and biochemical parameters, enzyme activities and serum cortisol in Besnoitia besnoiti naturally infected dairy cattle.

BACKGROUND: Besnoitia besnoiti is an Apicomplexan protozoa causative of bovine besnoitiosis, a chronic and debilitating disease of cattle, with a variety of pathological findings that could alter some laboratory parameters. A study was conducted in a bovine besnoitiosis endemically infected dairy herd located in Italy characterized by high intra-herd seroprevalence and cattle with clinical signs of the disease. In the study, alterations in laboratory parameters, i.e. hematological and biochemical parameters, enzyme activities and serum cortisol levels, in Besnoitia besnoiti naturally infected cows were investigated in depth. METHODS: Laboratory parameters in 107 cows, of which 61 were seronegative and 46 were seropositive to B. besnoiti, including 27 with clinical signs of bovine besnoitiosis, were compared. Generalized linear models were used to evaluate the effect of Besnoitia infection on the considered laboratory parameters. RESULTS: Hematological analyses revealed that B. besnoiti infection determined a significant alteration to the leukocyte differential, with a higher percentage of granulocytes and a lower percentage of lymphocytes in seropositive and clinically affected animals (Mann-Whitney U-test, P = 0.022); erythrocyte and platelet counts did not show any difference between the considered groups of cows. Biochemistry tests evidenced that the parasite infection influenced serum protein values in seropositive cows and glutamate dehydrogenase values in clinically affected animals. No or only slight differences were revealed for all of the other biochemical and enzyme activity parameters in B. besnoiti-infected animals. In addition, despite the lack of statistical significance, seropositive and clinically affected cows evidenced higher concentrations of serum cortisol values compared to seronegative animals. CONCLUSIONS: Although physiological, pathological and farm-related factors could have influenced the results in investigated animals, further studies involving more animals from different farms would be advisable to infer the role of B. besnoiti on these alterations, since laboratory parameters could help veterinarians in the diagnosis of bovine besnoitiosis in cattle.

Serological evaluation of Neospora caninum in pregnant women treated at referral center for prenatal screening in Mato Grosso do Sul, Brazil.

Neospora caninum is an obligate intracellular protozoan with canids (Canis domesticus, Canis lupus dingo, Canis latrans, Canis lupus) as its definitive hosts. The objective of this study was to detect anti-N. caninum antibodies in pregnant women seen at referral center for prenatal screening in the state of state Mato Grosso do Sul, Brazil. A total of 188 serum samples from pregnant women provided by the Instituto de Pesquisa, Ensino e Diagnósticos da APAE de Campo Grande (IPED/ APAE) were subjected to IFA test and western blot analysis. The samples were divided into three groups: 23/99 samples from the seropositive group for toxoplasmosis were positive for anti-N. caninum IgG antibodies, and 9/99 positive for IgM; in the HIV group, 7/33 were positive for IgG; and in the HIV+toxoplasmosis group, 13/56 were positive for IgG and two positive for IgM. The seropositivity for IgG was assessed by western blot by testing 43 IFA test positive samples using rNcSRS2 (Nc-p43) as antigen. The serological results of the present study suggest that exposure of these pregnant women to the parasite N. caninum and presence of IgM antibodies are indicative of recent infection. Further studies are needed to establish the possibility of active infection.

Seroprevalence and incidence of Toxoplasma gondii and Neospora caninum infection in naturally exposed domestic dogs from a rural area of São Paulo state, Brazil.

Dogs are hosts of the protozoans Toxoplasma gondii, which causes an important public health disease, and Neospora caninum. Studies that have evaluated toxoplasmosis and neosporosis for prolonged periods in dog populations are rare. We analyzed infection by both parasites in a domestic dog population over three consecutive years in São Paulo state, Brazil. In the 1st, 2nd and 3rd years of collection, 181, 193 and 172 domiciles were visited, and blood samples of 331, 371 and 348 dogs were collected for antibody serology, respectively. The seroprevalence of T. gondii in each year was 27.2%, 22.5% and 43.9%, respectively, and that of N. caninum was 7.8%, 4.8% and 6.8%, respectively. The incidence rates for T. gondii in the 2nd and 3rd collections were 13.2% and 30.0%, and those for N. caninum were 3.3% and 4.4%, respectively. Positive and negative serological conversions for both agents occurred at high frequencies during the study period. This study reveals the canine population's serological profile and demonstrates the constant exposure of dogs to the investigated pathogens, indicating the need for prevention and control measures in the region.

Influence of Eimeria spp. infection and dietary inclusion of arginine on intestine histological parameters, serum amino acid profile and ileal amino acids digestibility in broiler chicks.

Coccidiosis is considered to be one of the most important challenge in the poultry industry causes economic losses due to the destruction in the digestive tract of chicken. It disturbs amino acids profile and their digestibility, leading to weight lost and economic burden. Using dietary arginine may decrease the adverse effects of coccidiosis on chicken digestive tract. This study aimed to evaluate the effects of dietary inclusion of arginine on intestine histological parameters, serum amino acid concentration and ileal amino acid digestibility of broiler chicks infected with coccidiosis. A total number of 384 one-d-old broiler chicks (Ross 308) of mixed sex with initial weight of 42 ±â€¯2 g was allocated into 8 groups with 8 birds/pen from grower period. At 21 days of age, broiler chicks were infected with a mixture of Eimeria spp. Broiler chicks were divided into infected and un-infected groups and received arginine at recommended levels of 85, 100, 125 and 150 %. Intestinal morphology and lesions, serum amino acid concentration and ileal amino acid digestibility were evaluated. Broiler chicks infected with Eimeria spp. showed lower villus height and villus height: crypt depth ratio and also higher intestinal lesions (P < 0.05). Coccidia infection decreased the ileal amino acid digestibility for all studied amino acids and also reduced serum concentrations of amino acids, except lysine and isoleucine (P < 0.05). Dietary supplementation of arginine especially in higher levels significantly increased villus height and villus height:crypt depth ratio and decreased lesions (P < 0.05). Moreover, dietary supplementing of arginine increased the serum concentration of arginine (P < 0.05), but it did not have any significant effect on its digestibility (P > 0.05). In sum, coccidiosis decreases amino acid digestibility and serum amino acid concentration, but dietary inclusion of higher levels of arginine significantly improved histological parameters of broiler chicks infected with coccidiosis.

Endogenous transplacental transmission of Neospora caninum in successive generations of congenitally infected goats.

Neospora caninum is a protozoan that is considered an important agent of reproductive disorders in ruminants worldwide, and vertical transmission is the main form of infection and maintenance of neosporosis in herds. In goats, there have been no studies that have evaluated the transmission of N. caninum between successive generations. Thus, the objective of this study was to evaluate, through IFAT and PCR, the endogenous transplacental transmission of N. caninum in up to five generations of six families of dairy goats naturally infected by the parasite and whether it was possible for dairy goats to become free of infection over successive generations. Ninety-five serum samples from positive animals and 75 samples from negative animals were analyzed for N. caninum. Of the 95 samples analyzed, 93 contained anti-N. caninum antibodies (97.8 %). Titers of anti-N. caninum antibodies varied (increasing or decreasing) in the offspring; however, with an increase in the number of the goat generations, the offspring tended to have lower titers (p = 0.021) at the day of birth. Reproductive disorders such as abortions, stillbirth or fetal retention occurred at a rate of 10.4 % and were not influenced by the mother's titer of anti-N. caninum antibodies at the day of parturition or abortion. The results showed that infection by N. caninum persists throughout generations in congenitally infected goats.

Effect of coccidial challenge and vaccination on the performance, veterinary postmortem scores, and blood biochemistry of broiler chickens.

A total of 960 male Cobb 500 broilers were used in a growth performance study to explore the effect of coccidial vaccination and/or coccidial challenge on blood biochemistry and veterinary postmortem metrics. Day-old chicks were randomly divided into one of the 4 experimental treatments. Treatments were arranged in a 2 × 2 factorial arrangement, with the factors being without or with vaccination (administered on day 1) or coccidial challenge (oral gavage on day 7). Growth performance was monitored on a weekly basis. Blood sample collection, as well as full veterinary necropsies, were carried out on days 6, 8, 13, 20, 27, and 34. Birds that did not receive the vaccination but were challenged with coccidiosis had higher feed conversion ratio, lower body weights, and higher mortality than the other experimental groups, and this effect was particularly evident from day 13 to day 20. Birds challenged with coccidiosis had lower plasma sodium and total carotenoid concentrations and higher potassium and globulin concentrations than nonchallenged birds. Significant interactions between age and experimental treatment for these blood parameters were observed, particularly on day 13. The necropsy results confirmed the effectiveness of the challenge and vaccination treatments, wherein birds that were challenged had higher coccidiosis scores on day 13 and day 27 than birds that were not challenged. These results demonstrate the potential for plasma sodium, potassium, total protein, total carbon dioxide, globulin, and carotenoid analysis for early diagnosis of coccidiosis in growing broiler chickens. Further work is necessary to establish whether the changes in blood biochemistry observed in the present study are transferable to alternative flocks of chicken and whether early diagnosis and intervention may mitigate performance losses associated with this disease.

Validation and field evaluation of a competitive inhibition ELISA based on the recombinant protein tSAG1 to detect anti-Neospora caninum antibodies in sheep and goats.

Neospora caninum is a protozoan parasite that causes abortion and reproductive failure in small ruminants. We validated and evaluated under field conditions a competitive inhibition ELISA based on the truncated SAG1 protein (tSAG1) from N. caninum for the detection of anti-N. caninum antibodies in sheep and goat flocks. The assay was validated using 80 positive and 142 negative serum samples from sheep and goats analyzed by IFAT and immunoblot (IB). ciELISAtSAG1 was then used to evaluate the prevalence of anti-N. caninum antibodies in 1449 goats from 143 flocks and 385 sheep from 40 flocks and compared to IFAT. The prevalence of anti-Toxoplasma gondii antibodies was evaluated by IFAT. The ciELISAtSAG1 cut-off was ≥ 36 percent inhibition, with a diagnostic sensitivity of 100.0 % (95 % CI = 95.4-100.0 %) and a diagnostic specificity of 98.6 % (95 % CI = 95.0-99.8 %) relative to the agreement between IFAT and IB. The field evaluation revealed a concordance between ciELISAtSAG1 and IFAT of 97.4 %, with an agreement (κ) of 0.90 for sheep sera, and a concordance of 96.5 % with κ = 0.85 for goat sera. The overall prevalence of anti-N. caninum antibodies in sheep was 14.3 % by IFAT and 15.8 % by ciELISAtSAG1. In goats, prevalence was 12.9 % by IFAT and 14.6 % by ciELISAtSAG1. The overall prevalence of anti-T. gondii antibodies was 28.8 % in goats and 43.8 % in sheep. The ciELISAtSAG1 could be useful for large-scale detection of anti-N. caninum antibodies in sheep and goats, and for seroepidemiological investigations due to its appropriate sensitivity and specificity, and the simplicity of production.

Serological and molecular detection of Neospora caninum and Toxoplasma gondii in human umbilical cord blood and placental tissue samples.

Neosporosis primarily affects cattle and dogs and is not currently considered a zoonotic disease. Toxoplasmosis is a zoonosis with a worldwide distribution that is asymptomatic in most cases, but when acquired during pregnancy, it can have serious consequences. The seropositivity rates determined by the indirect fluorescent antibody test for Neospora caninum (N. caninum) and Toxoplasma gondii (T. gondii) were 24.3% (49 samples) and 26.8% (54 samples), respectively. PCR positivity for N. caninum was observed in two samples of cord blood (1%) using the Nc5 and ITS1 gene, positivity for T. gondii was observed in 16 samples using the primer for the B1 gene (5.5% positivity in cord blood and 2.5% positivity in placental tissue). None of the samples showed structures characteristic of tissue cysts or inflammatory infiltrate on histopathology. Significant associations were observed only between N. caninum seropositivity and the presence of domestic animals (p = 0.039) and presence of dogs (p = 0.038) and between T. gondii seropositivity and basic sanitation (p = 0.04). This study obtained important findings regarding the seroprevalence and molecular detection of N. caninum and T. gondii in pregnant women; however, more studies are necessary to establish a correlation between risk factors and infection.

Clinic Congenital Neosporosis in a Calf.

Neosporosis is an infectious disease which is caused by a protozoan called Neospora caninum and characterized by endemic and epidemic abortions in adult cattle and congenital encephalomyelitis in calves. Our case was a female calf from the Holstein breed which was born in a farm with abortion and infertility problems and was unable to stand up after birth.. It was stated that it was the third pregnancy of the mother cow and that the previous pregnancy was aborted in the fifth month. In clinical examination, sucking reflex of the calf was present and there was spastic paralysis starting from pelvic muscle in rear limbs. Blood (EDTA and spare tube) sample was taken from the cow and from the calf before sucking first colostrum and cerebrospinal fluid was collected from the calf.. Based on the clinical and serological findings, a diagnosis of congenital neosporosis was made. As a result, neosporosis must be considered in the clinical diagnosis of calves with neurological symptoms during the neonatal period.

Simultaneous differential detection of canine blood parasites: Multiplex high-resolution melting analysis (mHRM).

Recently, the incidence of canine infection by the tick-borne parasites Babesia spp., Hepatozoon canis, Ehrlichia canis and Anaplasma platys has been increasing globally. We have developed a multiplex high-resolution melting analysis (mHRM) technique to reduce the time demands and costs associated with detecting haemoparasites in canine blood, while increasing the degree of reliability of this method of analysis. We have designed primers that are specific for protozoans (B. vogeli and H. canis) and Rickettsia-like bacteria (E. canis and A. platys) based on the 18S or 16S rDNA sequences, respectively. Two primer pairs (Protz18S-C and Bact16S-A) were found to be suitable for detecting these agents since their melting temperatures (Tm) exhibited discernible differences among the four haemoparasites, A. platys, B. vogeli, E. canis and H. canis (83.10 °C, 82.41 °C, 80.37 °C and 78.56 °C, respectively). The sequences acquired from these PCR products were >94 % identical to those of A. platys, B. vogeli, E. canis and H. canis in GenBank. The limit of detection (LOD) for B. vogeli, E. canis and A. platys was 103 copies/µl, while the LOD for H. canis was 104 copies/µl. Of the 68 dogs tested, 28 (41 %) were infected with these agents. The most commonly occurring infection involved E. canis, followed by B. vogeli, A. platys and H. canis, with infection percentages of 26 %, 13 %, 7 % and 6 %, respectively. These results demonstrate that mHRM can serve as a rapid, economical and reliable tool for the detection of parasitic diseases in canine blood for diagnosis and epidemiology.

Species-specific differences in Toxoplasma gondii, Neospora caninum and Besnoitia besnoiti seroprevalence in Namibian wildlife.

BACKGROUND: Knowledge about parasitic infections is crucial information for animal health, particularly of free-ranging species that might come into contact with livestock and humans. METHODS: We investigated the seroprevalence of three tissue-cyst-forming apicomplexan parasites (Toxoplasma gondii, Neospora caninum and Besnoitia besnoiti) in 506 individuals of 12 wildlife species in Namibia using in-house enzyme linked immunosorbent assays (indirect ELISAs applying purified antigens) for screening and immunoblots as confirmatory tests. We included six species of the suborder Feliformia, four species of the suborder Caniformia and two species of the suborder Ruminantia. For the two species for which we had most samples and life-history information, i.e. cheetahs (Acinonyx jubatus, n = 250) and leopards (Panthera pardus, n = 58), we investigated T. gondii seroprevalence in relation to age class, sex, sociality (solitary, mother-offspring group, independent sibling group, coalition group) and site (natural habitat vs farmland). RESULTS: All but one carnivore species (bat-eared fox Otocyon megalotis, n = 4) were seropositive to T. gondii, with a seroprevalence ranging from 52.4% (131/250) in cheetahs to 93.2% (55/59) in African lions (Panthera leo). We also detected antibodies to T. gondii in 10.0% (2/20) of blue wildebeest (Connochaetes taurinus). Adult cheetahs and leopards were more likely to be seropositive to T. gondii than subadult conspecifics, whereas seroprevalence did not vary with sex, sociality and site. Furthermore, we measured antibodies to N. caninum in 15.4% (2/13) of brown hyenas (Hyaena brunnea) and 2.6% (1/39) of black-backed jackals (Canis mesomelas). Antibodies to B. besnoiti were detected in 3.4% (2/59) of African lions and 20.0% (4/20) of blue wildebeest. CONCLUSIONS: Our results demonstrate that Namibian wildlife species were exposed to apicomplexan parasites at different prevalences, depending on parasite and host species. In addition to serological work, molecular work is also needed to better understand the sylvatic cycle and the clear role of wildlife in the epidemiology of these parasites in southern Africa.

Seroprevalence and incidence of Toxoplasma gondii and Neospora caninum infection in naturally exposed domestic dogs from a rural area of São Paulo state, Brazil / Soroprevalência e incidência de infecção por Toxoplasma gondii e Neospora caninum em cães domésticos naturalmente expostos em uma área rural no estado de São Paulo, Brasil

Abstract Dogs are hosts of the protozoans Toxoplasma gondii, which causes an important public health disease, and Neospora caninum. Studies that have evaluated toxoplasmosis and neosporosis for prolonged periods in dog populations are rare. We analyzed infection by both parasites in a domestic dog population over three consecutive years in São Paulo state, Brazil. In the 1st, 2nd and 3rd years of collection, 181, 193 and 172 domiciles were visited, and blood samples of 331, 371 and 348 dogs were collected for antibody serology, respectively. The seroprevalence of T. gondii in each year was 27.2%, 22.5% and 43.9%, respectively, and that of N. caninum was 7.8%, 4.8% and 6.8%, respectively. The incidence rates for T. gondii in the 2nd and 3rd collections were 13.2% and 30.0%, and those for N. caninum were 3.3% and 4.4%, respectively. Positive and negative serological conversions for both agents occurred at high frequencies during the study period. This study reveals the canine population's serological profile and demonstrates the constant exposure of dogs to the investigated pathogens, indicating the need for prevention and control measures in the region.

Resumo Os cães são hospedeiros dos protozoários Toxoplasma gondii, que causam uma importante doença para a saúde pública, e Neospora caninum. Estudos que avaliam a toxoplasmose e a neosporose por períodos prolongados em populações caninas são raros. Foi analisada a infecção por esses dois parasitas em uma população de cães domésticos e domiciliados por três anos consecutivos, no Estado de São Paulo, Brasil. Nos 1º, 2º e 3º anos de coletas, 181, 193 e 172 domicílios foram visitados, nos quais foram coletadas amostras de sangue de 331, 371 e 348 cães para sorologia, respectivamente. A soroprevalência de T. gondii em cada ano foi de 27,2%, 22,5% e 43,9%, respectivamente; e a de N. caninum foi de 7,8%, 4,8% e 6,8%, respectivamente. As taxas de incidência para T. gondii, nas 2ª e 3ª coletas, foram de 13,2% e 30,0%, respectivamente; e para N. caninum, 3,3% e 4,4%, respectivamente. As conversões sorológicas positivas e negativas para ambos os agentes ocorreram em alta frequência durante o período analisado. Este estudo revela o perfil sorológico da população canina e demonstra sua constante exposição aos patógenos investigados, o que requer medidas de prevenção e controle na região.

Serological evaluation of Neospora caninum in pregnant women treated at referral center for prenatal screening in Mato Grosso do Sul, Brazil / Avaliação sorológica de Neospora caninum em gestantes atendidas em centro de referência para triagem pré-natal em Mato Grosso do Sul, Brasil

Abstract Neospora caninum is an obligate intracellular protozoan with canids (Canis domesticus, Canis lupus dingo, Canis latrans, Canis lupus) as its definitive hosts. The objective of this study was to detect anti-N. caninum antibodies in pregnant women seen at referral center for prenatal screening in the state of state Mato Grosso do Sul, Brazil. A total of 188 serum samples from pregnant women provided by the Instituto de Pesquisa, Ensino e Diagnósticos da APAE de Campo Grande (IPED/ APAE) were subjected to IFA test and western blot analysis. The samples were divided into three groups: 23/99 samples from the seropositive group for toxoplasmosis were positive for anti-N. caninum IgG antibodies, and 9/99 positive for IgM; in the HIV group, 7/33 were positive for IgG; and in the HIV+toxoplasmosis group, 13/56 were positive for IgG and two positive for IgM. The seropositivity for IgG was assessed by western blot by testing 43 IFA test positive samples using rNcSRS2 (Nc-p43) as antigen. The serological results of the present study suggest that exposure of these pregnant women to the parasite N. caninum and presence of IgM antibodies are indicative of recent infection. Further studies are needed to establish the possibility of active infection.

Resumo Neospora caninum é um protozoário intracelular obrigatório que possui os canídeos (Canis domesticus, Canis lupus dingo, Canis latrans, Canis lupus) como seus hospedeiros definitivos. O objetivo deste trabalho foi detectar anticorpos anti-N. caninum em gestantes, atendidas em centro de referência para triagem pré-natal, em Mato Grosso do Sul, Brasil. Um total de 188 amostras de soro de gestantes, cedidas pelo Instituto de Pesquisa, Ensino e Diagnósticos da APAE de Campo Grande (IPED/ APAE), foram submetidas ao teste de RIFI e western blot para a detecção de anticorpos anti-N. caninum. As amostras foram divididas em três grupos: 23/99 amostras do grupo soropositivo para toxoplasmose demonstraram positividade para anticorpos IgG anti-N. caninum e 9/99 positivos para IgM. No grupo HIV 7/33 apresentaram positividade para IgG. No grupo HIV+Toxoplasmose 13/56 apresentaram positividade para IgG e duas para IgM. A soropositividade para IgG foi avaliada por western blot, testando 43 amostras positivas para RIFI, usando-se rNcSRS2 (Nc-p43) como antígeno. Os resultados sorológicos do presente estudo sugerem exposição dessas gestantes ao parasita N. caninum e a positividade para anticorpos IgM são indicativos de infecção recente. Mais estudos na área são necessários para estabelecer a possibilidade de infecção ativa.

Diversity of Karyolysus and Schellackia from the Iberian lizard Lacerta schreiberi with sequence data from engorged ticks.

Apicomplexan haemoparasites of the genera Schellackia Reichenow, 1919, and Karyolysus Labbé, 1894, seem to be common in lizards and widespread across the world. For decades, their identification has been based on morphological descriptions and life cycle patterns, with molecular characterizations, applied only recently. We used molecular characterization to confirm the identification of haemoparasites detected by microscopy in blood smears of Lacerta schreiberi Bedriaga, 1878, a lizard of the Iberian Peninsula. Since blood samples other than blood smears were not available from the studied lizards, 264 engorged ticks Ixodes ricinus (Linneaus, 1758) collected from them were used as an alternative non-invasive source of haemoparasite DNA for molecular genetic analyses. Of the 48 blood smears microscopically examined, 31 were positive for blood parasites (64.6% prevalence). We identified trophozoites and gamonts similar to Karyolysus lacazei (Labbé, 1894) (24/48; 50%) and Schellackia-like sporozoites (20/48; 41.7%). Mixed infections with both species occurred in 13 blood smears (27.1%). Sequence data were obtained for both parasites from engorged ticks. Phylogenetic analyses placed our unique haemogregarine sequence within the Karyolysus clade, nevertheless, within substantial polytomy. Thus, according to its morphology and effect on the host cell, we refer to this haemogregarine as Karyolysus cf. lacazei. Besides the Schellackia sequences being identical to a previously identified haplotype, we also obtained sequences of three new closely related haplotypes.