Effectiveness of Heterologous Coronavirus Disease 2019 (COVID-19) Vaccine Booster Dosing in Brazilian Healthcare Workers, 2021.

BACKGROUND: Little is currently known about vaccine effectiveness (VE) for either 2 doses of Oxford-AstraZeneca (ChAdOx1) viral vector vaccine or CoronaVac (Instituto Butantan) inactivated viral vaccine followed by a third dose of mRNA vaccine (Pfizer/BioNTech) among healthcare workers (HCWs). METHODS: We conducted a retrospective cohort study among HCWs (aged ≥18 years) working in a private healthcare system in Brazil from January to December 2021. VE was defined as 1 - incidence rate ratio (IRR), with IRR determined using Poisson models with the occurrence of laboratory-confirmed coronavirus disease 2019 (COVID-19) infection as the outcome, adjusting for age, sex, and job type. We compared those receiving viral vector or inactivated viral primary series (2 doses) with those who received an mRNA booster. RESULTS: A total of 11 427 HCWs met the inclusion criteria. COVID-19 was confirmed in 31.5% of HCWs receiving 2 doses of CoronaVac vaccine versus 0.9% of HCWs receiving 2 doses of CoronaVac vaccine with mRNA booster (P < .001) and 9.8% of HCWs receiving 2 doses of ChAdOx1 vaccine versus 1% among HCWs receiving 2 doses of ChAdOx1 vaccine with mRNA booster (P < .001). In the adjusted analyses, the estimated VE was 92.0% for 2 CoronaVac vaccines plus mRNA booster and 60.2% for 2 ChAdOx1 vaccines plus mRNA booster, when compared with those with no mRNA booster. Of 246 samples screened for mutations, 191 (77.6%) were Delta variants. CONCLUSIONS: While 2 doses of ChAdOx1 or CoronaVac vaccines prevent COVID-19, the addition of a Pfizer/BioNTech booster provided significantly more protection.

The Hip of Children with Congenital Zika Syndrome: A Prospective Observational Study.

OBJECTIVE: To assess the clinical and radiographic characteristics of hip joint deformities in children with congenital Zika syndrome (CZS), and the evolution of hip joint deformities in affected infants for the first 3 years of life. STUDY DESIGN: This prospective observational study evaluated orthopedic clinical examinations performed every 3 months to assess hip flexion and extension, lateral and medial rotation, and abduction and adduction, as well as lower limb muscle length and tone. The biannual radiograph comprised anteroposterior panoramic pelvic radiographs with the lower limbs in extension. Percentage of migration was used as a radiographic study tool to measure and evaluate linear hip displacement. RESULTS: From November 2018 to March 2020, we followed 30 children with CZS, of whom 15 (50%) had normal pelvic radiographs on admission; 5 (33.3%) developed hip displacement by the second radiograph examination. During follow-up radiographic examinations, 20 of the 30 children (66.7%) were diagnosed with hip displacement and/or dislocation of at least 1 side, and 10 of the 30 (33.3%) remained normal. Among 30 affected patients, 13 (43.3%) had hip displacement on the right side and 9 (30%) on the left side. Logistic regression analysis revealed that spasticity (P = .0033; OR, 15.9) and ophthalmologic abnormalities (P = .0163; OR, 16.9) were associated with hip dislocation during follow-up. CONCLUSIONS: Pelvic radiographic follow-up for all children with CZS will complement physical examination, diagnosis, and monitoring for hip joint deformities.

Evaluation and monitoring of eye findings in children exposed to Zika virus during gestation: 3 years of follow-up.

INTRODUCTION: Congenital Zika syndrome (CZS) is a recently described disease. Our main objective was to evaluate and monitor, over 3 years, the ophthalmoscopic findings in children exposed to zika virus (ZIKV) during gestation. METHODS: This prospective observational study was conducted in Rio de Janeiro, Brazil, between April 2016 and May 2019. We evaluated two groups with exanthema serving as a proxy for viremia: (i) children whose mothers had exanthema during pregnancy and (ii) children who had microcephaly without maternal exanthema during pregnancy. We performed indirect ophthalmoscopy at recruitment and every 6 months thereafter. We also tested the association between ocular findings with maternal exanthema, microcephaly, CZS and maternal infection confirmed by reverse transcriptase quantitative polymerase chain reaction and gender. RESULTS: Of the 72 children included, 16 (22.2%) had optic nerve and/or retinal lesions. All 16 had CZS and 15 (93.7%) had microcephaly (14 at birth and 1 postnatally). The child with postnatally acquired microcephaly was born to a mother without exanthema during pregnancy. Fifty-six (77.8%) of the 72 children were followed for a median time of 24 months and none exhibited differences between admission and follow-up examinations. After logistic regression, only microcephaly at birth was associated with eye abnormalities (odds ratio, 77.015; 95% confidence interval, 8.85-670.38; p < 0.001). CONCLUSION: We observed that there was no progression of the lesions over the follow-up period. We also showed that the eye findings were associated only with microcephaly at birth. Attention should be paid to all children born during a ZIKV epidemic, regardless of maternal exanthema and/or microcephaly at birth.

Two-year follow-up of children with congenital Zika syndrome: the evolution of clinical patterns.

The aim of the study was to describe neurological manifestations in children with congenital Zika syndrome (CZS) in the first 2 years of age. In this prospective observational study, children with CZS treated at a university hospital received a neurological assessment and were evaluated using two neurodevelopmental scales (the Denver II test and the assessment of gross motor development of the World Health Organization) by a pediatric neurologist on admission to the study and at 4, 8, 12, 18, and 24 months of age. The data collected were stored in Microsoft Excel version 14.6.3. Thirty-eight children (27 males and 11 females; a median age of 4.3 months (interquartile range (IQR): 1.6-11.4)) with CZS were evaluated. Irritability was present in 50% and 27% of the children at 8 months and 24 months, respectively. Axial hypertonia was highly prevalent at 4 months (77%), with a decrease to 50% at 24 months. At all ages, spastic tetraparesis was the most common motor abnormality (> 80%). Twenty-seven (71%) participants were diagnosed with epilepsy, and the median age at seizure onset was 6 months (IQR: 3.5-8). The most frequent types of seizures were focal seizures and spasms, with spasms being the most frequent in the first year of life (52%) and focal crises being the most frequent in the second year of life (50%).Conclusion: This study allowed observation of neurological abnormalities over time, the evolution of epileptic manifestations, and recognition of new patterns of clinical neurological abnormalities, helping clinicians to recognize CZS earlier, minimizing the impact of new outbreaks. What is Known: • Clinical patterns of SZC patients at pre-established ages or date of data collection • More frequent studies with data collection of clinical-radiological features of patient's over his first year of life What is New: • Comprehensive clinical neurological progression data regarding CZS in the first 2 years of life, recognizing patterns • Hypothesis including a new CZS spectrum with milder clinical-radiological features.

Validity and reliability of the Brazilian version of the Body Image in Pregnancy Scale.

OBJECTIVES: To describe the translation and cultural adaptation, and evaluate the psychometric properties of the Body Image in Pregnancy Scale when applied to Brazilian pregnant women. BACKGROUND: Poor body image of pregnant women is associated with negative outcomes for maternal and child health. However, there is a lack of psychometric studies aiming to evaluate the body image experiences during pregnancy. METHODS: A methodological study was carried out with 187 Brazilian pregnant women. We performed translation and cross-cultural adaptation (evidence of content validity), and evaluated the psychometric properties (factorial validity through principal component analysis, discriminant validity, and estimated internal consistency) of the Brazilian version of the Body Image in Pregnancy Scale. RESULTS: The scale items presented semantic, idiomatic, cultural, and conceptual equivalences, showing evidence of content validity. Principal component analysis resulted in a 6-factor structure with 35 items which showed an adequate adjustment of the scale among Brazilian pregnant women. Adequate estimated internal consistency (Cronbach's α = .88) was found. The Brazilian version was not able to discriminate pregnant women from different gestational age. CONCLUSION: The Brazilian version of the Body Image in Pregnancy Scale showed evidence of content validity, factorial validity, and estimated internal consistency.

The impact of dry ageing vacuum-packed pork on the viability of Toxoplasma gondii tissue cysts.

The present study evaluated the viability of Toxoplasma gondii tissue cysts in dry-aged pork loins (m. longissimus) after 14, 21 and 28 days under controlled temperature (0 °C ±â€¯1 °C). The pigs (n = 9) were orally inoculated with 3,000 T. gondii oocysts. The right loin of each pig was aged for a predetermined period, and the left loin was kept unprocessed as a control. Two experiments were performed. In Experiment 1, the loins of three pigs were aged for 14 days and then bioassayed in both cats and mice. In Experiment 2, the loins of six pigs were bioassayed only in mice, and the ageing periods were 14, 21, and 28 days. Toxoplasma gondii tissue cysts remained viable in loins aged up to 14 days, as confirmed by bioassays in cats and mice. Viable T. gondii was not recovered by bioassays in mice from loins that were aged for 21 or 28 days. These results demonstrate that T. gondii remained viable in vacuum-packed dry-aged pork loins for 14 days at controlled temperature but not for 21 days or longer.

Isolation of viable Toxoplasma gondii from organs and Brazilian commercial meat cuts of experimentally infected pigs.

The present study evaluated the distribution and viability of Toxoplasma gondii tissue cysts in the organs and Brazilian commercial cuts of experimentally infected pigs. The pigs were infected with 3 × 103 oocysts of the T. gondii isolate TgCkBr57 (Type BrII). Mouse bioassays were performed on the brain, retina, tongue, diaphragm, and heart as well as the following muscle cuts: loin (longissimus), coppa (longissimus, spinalis dorsi, rhomboideus), tenderloin (psoas major), outside flat (biceps femoris), topside (semimembranosus), and top sirloin (gluteus medius). Toxoplasma gondii was isolated from the coppa, heart, diaphragm, and tongue of three pigs; from the tenderloin, outside flat, and brain of two pigs; and from the top sirloin and loin of one pig. Thus, the viability of T. gondii cysts was observed in all of the organs and cuts evaluated (except for the topside and retina), demonstrating the broad distribution of this parasite in pig organs and commercial meat cuts, and the importance of this species as a source of human infection.

Outbreak of toxoplasmosis in a flock of domestic chickens (Gallus gallus domesticus) and guinea fowl (Numida meleagris).

Toxoplasmosis is a disease with a worldwide distribution that affects a wide variety of animal species, though with rare descriptions in chickens. We describe the clinical, epidemiological, pathological, and molecular aspects of a toxoplasmosis outbreak in domestic chickens and guinea fowl in southern Brazil. The flock was composed of 47 domestic chickens and 29 guinea fowl. Of these, 22 birds showed clinical signs of lethargy, anorexia, and neurological signs over a clinical course of 24-72 h, and 15 died. Epidemiological data were obtained through fieldwork performed at the chicken farm and necropsies of six birds. Gross lesions were absent at necropsy, and histopathological findings included inflammatory infiltrate of macrophages, lymphocytes, and plasma cells and necrosis in several tissues associated with intralesional Toxoplasma gondii. Immunohistochemistry for T. gondii was positive. Additionally, restriction fragment length polymorphism (RFLP) analysis with 11 markers (SAG1, SAG2 (5'3'SAG2 and alt. SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3) and microsatellite (MS) analysis with 15 markers (TUB2, W35, TgMA, B18, B17, M33, IV.1, XI.1, N60, N82, AA, N61, N83, M48, and M102) were performed. PCR-RFLP revealed T. gondii genotype ToxoDB-PCR-RFLP #280, and MS analysis also showed a unique genotype. This is the first description of this genotype in chickens and adds to the evidence suggesting considerable genotypic diversity of T. gondii in Brazil.

Children Born to Mothers with Rash During Zika Virus Epidemic in Brazil: First 18 Months of Life.

OBJECTIVE: To better understand the clinical spectrum and course of congenital Zika syndrome (CZS) during the first 18 months of life of children whose mothers had rash during pregnancy. METHODS: This longitudinal observational study evaluated the clinical progress from birth until 18 months of life of children of mothers who developed rash during or up to 3 months before gestation. Maternal rash occurred from November 2015 to May 2017. The study subjects were divided into three groups: children whose mothers tested positive by RT-qPCR for Zika virus (ZIKV) (Group 1), children whose mothers tested negative by RT-qPCR for ZIKV (Group 2), and children whose mothers did not undergo any testing for ZIKV (Group 3) but tested negative for other congenital infections. RESULTS: Between April 2016 and July 2018, we studied 108 children: 43 in Group 1, 26 in Group 2 and 39 in Group 3. The majority of children were admitted into the study within 6 months of life. CZS was diagnosed in 26 children, equally distributed in Groups 1 and 3. Of 18 children with microcephaly, 6 were in Group 1 (1 postnatal) and 12 were in Group 3 (5 postnatal). Maternal rash frequency was 10 times higher during the first trimester than in the other trimesters (OR: 10.35; CI 95%: 3.52-30.41). CZS was diagnosed during the follow-up period in 14 (54%) cases. Developmental delays and motor abnormalities occurred in all children and persisted up to 18 months. Epilepsy occurred in 18 (69%) of the cases. CONCLUSIONS: Infants born of mothers exposed to ZIKV during pregnancy showed progression of developmental, motor and neurologic abnormalities even if they were born asymptomatic. Continued postnatal monitoring of such newborns is necessary to preclude disability-associated complications.

Typical Brazilian genotype of Toxoplasma gondii isolated from a horse destined for human consumption in Europe from a slaughterhouse.

Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii. Infections occur via the ingestion of oocysts, consumption of cysts containing bradyzoites, and transplacental transmission of tachyzoites. Diversity in T. gondii strains may affect the outcome of clinical toxoplasmosis. The consumption of horse meat is a common practice in some parts of the world. The objectives of the present study were to isolate and genotype T. gondii from horses from an abattoir in the state of Rio Grande do Sul in southern Brazil that exports horse meat to Europe. Antibodies to T. gondii were found in 32.5% (13/40) of the horses using the modified agglutination test (MAT) with a cut-off of 1:25. Tissues from the 13 seropositive horses were bioassayed in mice, and one isolate, designated TgHorseBrRS1, was obtained. PCR-RFLP of the isolate revealed the ToxoDB-RFLP #228 genotype, a typical non-archetypal Brazilian genotype, and microsatellite analysis showed a unique non-archetypal genotype. This study showed that horses from Brazil can harbor viable T. gondii in their tissues, suggesting that recommendations to consumers should be made, especially in European countries where consumption of raw horse meat is common.

Survey of Plant Growth-Promoting Mechanisms in Native Portuguese Chickpea Mesorhizobium Isolates.

Rhizobia may possess other plant growth-promoting mechanisms besides nitrogen fixation. These mechanisms and the tolerance to different environmental factors, such as metals, may contribute to the use of rhizobia inocula to establish a successful legume-rhizobia symbiosis. Our goal was to characterize a collection of native Portuguese chickpea Mesorhizobium isolates in terms of plant growth-promoting (PGP) traits and tolerance to different metals as well as to investigate whether these characteristics are related to the biogeography of the isolates. The occurrence of six PGP mechanisms and tolerance to five metals were evaluated in 61 chickpea Mesorhizobium isolates previously obtained from distinct provinces in Portugal and assigned to different species clusters. Chickpea microsymbionts show high diversity in terms of PGP traits as well as in their ability to tolerate different metals. All isolates synthesized indoleacetic acid, 50 isolates produced siderophores, 19 isolates solubilized phosphate, 12 isolates displayed acid phosphatase activity, and 22 exhibited cytokinin activity. Most isolates tolerated Zn or Pb but not Ni, Co, or Cu. Several associations between specific PGP mechanisms and the province of origin and species clusters of the isolates were found. Our data suggests that the isolate's tolerance to metals and ability to solubilize inorganic phosphate and to produce IAA may be responsible for the persistence and distribution of the native Portuguese chickpea Mesorhizobium species. Furthermore, this study revealed several chickpea microsymbionts with potential as PGP rhizobacteria as well as for utilization in phytoremediation strategies.

Isolation and biological and molecular characterization of Neospora caninum (NC-SP1) from a naturally infected adult asymptomatic cattle (Bos taurus) in the state of São Paulo, Brazil.

The biological and genetic diversity of Neospora caninum is very limited because of availability of only a few viable isolates worldwide. This study describes the isolation and biological and molecular characterization of a new viable isolate of N. caninum (NC-SP1), from a cattle in Brazil. Approximately 400 g of brain from a naturally infected adult male cattle from an abattoir was fed to a 2-month-old dog. Neospora-like oocysts were observed on day 7 post-inoculation (PI) and the duration of oocyst shedding was 14 days. The DNA obtained from oocysts was characterized molecularly and the final sequence was 99% identical to homologous sequences of N. caninum available in GenBank®. For bioassay, gerbils (Meriones unguiculatus) were orally inoculated with 10 100 and 1000 oocysts; all gerbils remained clinically normal but developed N. caninum antibodies 14 days PI. Cell culture isolation was successful using the brain homogenate from one of the gerbils and tachyzoites were observed 24 days PI. Microsatellite genotyping revealed a unique genetic profile for this new reference isolate.

Dengue Hemorrhagic Fever: A State-of-the-Art Review Focused in Pulmonary Involvement.

Dengue fever is an arboviral disease transmitted to humans through the bites of infected female Aedes mosquitoes. Dengue virus is a member of the Flaviviridae family, and human infection can be caused by any of the four antigenically distinct serotypes (DENV 1-4). The infection has become recognized as the most important and prevalent arboviral disease in humans, endemic in almost 100 countries worldwide. Nearly 3 billion people live in areas with transmission risk. Autochthonous transmission of the virus in previously disease-free areas, increased incidence in endemic areas, and epidemic resurgence in controlled regions could increase the risk of contracting more severe forms of the disease, such as dengue hemorrhagic fever (DHF)/dengue shock syndrome (DSS). Symptomatic dengue virus infection can present with a wide range of clinical manifestations, from mild fever to life-threatening DSS. Thoracic complications may manifest as pleural effusion, pneumonitis, non-cardiogenic pulmonary edema, and hemorrhage/hemoptysis. No vaccine is currently available and no specific treatment for dengue fever exists, but prevention and prompt management of complications in patients with DHF can help reduce mortality. This review describes the main clinical, pathological, and imaging findings of thoracic involvement in DHF.

Rare case of acute toxoplasmosis in a domestic rabbit (Oryctolagus cuniculus) in Brazil associated with the type BrIII Brazilian clonal lineage of Toxoplasma gondii.

Toxoplasmosis is a widely distributed disease that infects birds and mammals, including humans. Acute clinical course of toxoplasmosis is considered to be rare among domestic rabbits (Oryctolagus cuniculus). The aim of this study was to present the first report of fatal acute disease caused by Toxoplasma gondii type BrIII genotype, a typical Brazilian clonal lineage, in a domestic rabbit. T. gondii was identified in histological sections of spleen and liver tissue, and these tissues were also immunohistochemically positive for T. gondii. After the histopathological and immunohistochemical confirmation of T. gondii, the genotype of this pathogen was determined via PCR-RFLP with 11 markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3) and via microsatellite (MS) analysis with 15 markers (TUB2, W35, TgMA, B18, B17, M33, IV.1, X1.1, M48, M102, N60, N82, AA, N61, and N83). This study shows that type BrIII genotype, circulating in Brazil in different hosts, can cause acute disease in a naturally infected animal host. The described case also involves the first reported occurrence of the 291 allele for the typing marker TUB2 in a type BrIII strain, emphasizing the genetic diversity of T. gondii in Brazil.

Acute toxoplasmosis in pigs in Brazil caused by Toxoplasma gondii genotype Chinese 1.

This study reports the clinicopathological, immunohistochemical, and molecular findings from two cases of systemic toxoplasmosis in pigs showing apathy and dyspnea. In the post-mortem examination, severe diffuse necrotizing bronchointerstitial pneumonia with numerous intralesional tachyzoites of Toxoplasma gondii was observed. The lungs had not collapsed but were diffusely reddened, and the parenchyma showed friable whitish subpleural nodules with multifocal to coalescent distribution and diameters of 0.5-1.0 cm. The histopathological findings comprised mononuclear inflammation and multifocal areas of necrosis in alveolar septa (cases 1 and 2). In addition, esophagitis and ulcerations in the mucosa of the stomach and the small and large intestines were observed (case 1). Immunohistochemical analysis using anti-T. gondii antibodies on lung tissue in both cases revealed strong immunolabeling of free tachyzoites and tachyzoites in the cytoplasm of histiocytes and in cysts. Nested PCR targeting a 155-bp fragment of the B1 gene of T. gondii was positive for the DNA extracted from lung fragments from the two pigs. Genotyping of the samples by means of PCR-RFLP (10 markers) and by means of microsatellites (15 of them) revealed that these animals were infected with T. gondii that was molecularly characterized as the non-archetypal genotype Chinese 1. This presents worldwide circulation, but it had not previously been described in Brazil. The microsatellite analysis showed that the animals were infected with the same T. gondii isolate circulating in the environment.

Genes commonly involved in acid tolerance are not overexpressed in the plant microsymbiont Mesorhizobium loti MAFF303099 upon acidic shock.

Rhizobia are legume bacterial symbionts that fix nitrogen in the root nodules of plants. The aim of the present study was to investigate the global transcriptional response of rhizobia upon an acidic shock. Changes in the transcriptome of cells of Mesorhizobium loti strain MAFF303099 upon an acidic shock at pH 3 for 30 min were analysed. From a total of 7,231 protein-coding genes, 433 were found to be differentially expressed upon acidic shock, of which 322 were overexpressed. Although most of the overexpressed genes encode hypothetical proteins, the two most represented Cluster of Orthologous Group (COG) categories are 'defence mechanisms' and 'transcription'. Differentially expressed genes are dispersed throughout the chromosome, with the exception of the symbiosis island, where most genes remain unchanged. A significant number of transcriptional regulators and ABC transporter genes are overexpressed. No overexpression of genes typically associated to acid tolerance in rhizobia, such as act and exo genes, was detected. Overall, this study suggests a transcriptional response to acidic shock of M. loti distinct from other rhizobia. Additional studies are in course to explore the role of some of the highly overexpressed genes and to further elucidate the molecular bases of acid stress response.

Clinical features and laboratory findings of human parvovirus B19 in human immunodeficiency virus-infected patients.

Immunocompromised patients may develop severe chronic anaemia when infected by human parvovirus B19 (B19V). However, this is not the case in human immunodeficiency virus (HIV)-infected patients with good adherence to highly active antiretroviral treatment (HAART). In this study, we investigated the clinical evolution of five HIV-infected patients receiving HAART who had B19V infections confirmed by serum polymerase chain reaction. Four of the patients were infected with genotype 1a strains and the remaining patient was infected with a genotype 3b strain. Anaemia was detected in three of the patients, but all patients recovered without requiring immunoglobulin and/or blood transfusions. In all cases, the attending physicians did not suspect the B19V infections. There was no apparent relationship between the infecting genotype and the clinical course. In the HAART era, B19V infections in HIV-positive patients may be limited, subtle or unapparent.

Nutritional status of HIV-positive patients in Niterói, Rio de Janeiro, Brazil.

The objective of this cross-sectional study was to assess the nutritional status of HIV-positive patients in a hospital in the city of Niteroi, Rio de Janeiro, Brazil. We studied 235 patients (130 men and 105 women) from May 2009 to June 2010. The frequency of undernourishment among women was 7.6%; 26.7% of the women were overweight, and 16.2% were obese. Among men, the frequency of undernourishment was 3.8%; 25.4% of the men were overweight, and 6.9% were obese. A logistic regression was done to investigate the relationship between nutritional status and potential predisposing factors. Women were more frequently affected by obesity and undernourishment than men. However, only the difference in obesity was significant, and women had almost three times higher odds of being obese (OR 2.6; 95% CI 1.03-6.65). According to a nationwide survey done in Brazil during 2008-2009, 50.1% of the Brazilian healthy males were overweight, and 12.5% were obese; 48% of healthy females were overweight, and 16.9% were obese. Although the prevalence of undernourishment in HIV-positive patients is now lower than that observed in the beginning of the AIDS epidemic and excess weight is increasingly common among people living with HIV/AIDS, the proportion of excess weight was found lower and of undernourishment was higher in the present study than that found in the Brazilian population.

Isolation and genetic characterization of Toxoplasma gondii from alpaca (Vicugna pacos) and sheep (Ovis aries).

Alpacas are important to the economy of several countries. Little is known of Toxoplasma gondii infection in alpacas worldwide. In the present study, T. gondii was isolated and genetically characterized from alpacas for the first time. Alpacas (n = 16) and rams (n = 12) pastured on a farm in Virginia, USA, were examined at necropsy. Antibodies to T. gondii were determined by the modified agglutination test (MAT, 1:25) and found in 6 of 16 alpacas with titers of 1:100 (2 alpaca), 1:400 (2 alpacas), 1:800 (1 alpaca), and 1:1,600 (1 alpaca), and 5 of 12 rams in titers of 1:50 in one, 1:400 in one, 1:800 in one, 1:1,600 in one, and 1:3,200 in one. Tissues of all 16 alpacas were bioassayed in mice or in cats. Muscles (heart, skeletal muscle) of nine alpacas with MAT titers of 1:25 were fed to T. gondii-free cats; the cats did not shed oocysts. Viable T. gondii was isolated from tissues of two of six seropositive alpacas by bioassay in mice. Viable T. gondii was isolated from three of three seropositive sheep by bioassay in mice. Genotyping using cell-cultured tachyzoites revealed four genotypes, including one for ToxoDB PCR-RFLP genotype #2 (type III), one for genotype #3 (type II variant), one for genotype #170, and two for a new genotype designated as ToxoDB PCR-RFLP genotype #230. Thus, four of the five T. gondii isolates in the present study belonged to different genotypes. These results indicate a higher genetic diversity among T. gondii isolates circulating in the USA than previously realized.

Evidence of Natural Zika Virus Infection in Captive Cervid Species in Brazil.

As part of an epidemiologic study of the Zika virus (ZIKV) in deer (Xxxx), samples from 56 captive deer in south and southeastern Brazil were tested for evidence of ZIKV. Three samples were positive using reverse-transcription quantitative PCR, although no samples were positive by virus isolation.