Neospora caninum antibodies in bulk tank milk from dairy cattle herds in Italy in relation to reproductive and productive parameters and spatial analysis.

Among the available diagnostic techniques, antibody detection in bulk tank milk (BTM) represents a useful tool to estimate and monitor Neospora caninum herd prevalence. To evaluate the prevalence of N. caninum and the effect of parasite infection on herd performances, BTM samples collected from 586 dairy herds located in one of the largest dairy production areas in Italy (Lombardy) were analyzed by an indirect ELISA to detect anti-N. caninum specific antibodies. Generalized linear models (GLMs) were developed. A purely spatial analysis scanning for clusters with high or low rates for N. caninum using the Bernoulli model was performed. A maximum entropy approach was used to estimate the probability of distribution of the parasite based on occurrence records together with environmental variables. Overall, 180 herds resulted positive for N. caninum antibodies on bulk tank milk (P = 30.7 %). A higher risk of seropositivity was evidenced in the provinces of Milano, Cremona, Brescia, and Bergamo (P = 32-40 %); a lower risk was evidenced in Lodi, Pavia, and Mantova (P = 13-24 %). A higher risk of seropositivity was revealed for small-medium farms (101-300 animals) (O.R.=2.8) and for older animals with more than 4 years (O.R.=4.4). Regarding the effect of N. caninum infection on herd performances, the number of inseminations for conception was higher (> 3 inseminations), and the period from calving to conception was longer (> 150 days) for positive farms (O.R.=2.0 and O.R.=2.3, respectively); besides, lower head daily milk production (<20 kg and 21-25 kg) and mature equivalent milk yield (<11,000), and somatic cell counts higher than 300,000 cells/ml were observed for N. caninum positive herds (O.R.=0.4, O.R.=0.4 and O.R.=1.9 respectively). The geographical distribution of N. caninum positive farms with the highest level of probability covers the central sector of the Po Plain where a significant cluster for high risk of parasite infection was shown by spatial scan statistic and Maximum entropy ecological niche modelling. A further significant cluster of low risk occurred in the southern. The climatic and environmental variables with the highest training gain when used in isolation resulted altitude, land use/land cover, and other variables related to temperature and precipitation. Neosporosis is widely distributed in Italian dairy herds and an impact of the parasite on herd performances could be hypothesized. Even if the role of N. caninum in alterations of reproductive and productive parameters should be further explored, veterinarians and farmers should be aware of neosporosis, and control plans should be adopted.

First Report of Isolation and Characterization of Acanthamoeba spp. from the Milk Used for Calf Feeding.

PURPOSE: Acanthamoeba spp. can be found in natural and artificial environments, which reflects their high adaptability to different conditions. Based on the available data, there is scarce information about the isolation of amoeba from milk. This study aimed to investigate the probable presence of Acanthamoeba in milk used for calf feeding. METHODS: 200 milk samples from 50 industrial and traditional farms were collected. The samples were filtered and cultured on the 1.5% Non-nutrient agar medium. The amoebic growth was examined with an inverted microscope daily. DNA was extracted from the positive plates, and a PCR reaction was undertaken using the primers amplifying the Acanthamoeba 18 S rRNA gene. Five samples were purified and sequenced using specific primers. Maximum likelihood reconstructions were performed using the phylogenetic program MEGA software. The osmo and thermotolerance of isolated trophozoites were examined as well. RESULTS: Out of 200 milk samples, Acanthamoeba was isolated from 27 (13.5%). The phylogenetic tree represents that all the isolates belonged to the genotype T4. Results of thermo and osmotolerance tests showed that isolates could develop at 37 and 43 ◦C. Besides, trophozoites survived at 0.5 M mannitol and 1 M. CONCLUSION: For the first time, Acanthamoeba spp. were isolated from milk used to feed dairy calves. Due to Acanthamoeba's neglected role in pathogen persistence and survival, hygiene instructions should be reconsidered.

Multinomial logistic regression based on neural networks reveals inherent differences among dairy farms depending on the differential exposure to Fasciola hepatica and Ostertagia ostertagi.

Fasciola hepatica and Ostertagia ostertagi are cattle parasites with worldwide relevance for economic outcome as well as animal health and welfare. The on-farm exposure of cattle to both parasites is a function of host-associated, intrinsic, as well as environmental and farm-specific, extrinsic, factors. Even though knowledge on the biology of both parasites exists, sophisticated and innovative modelling approaches can help to deepen our understanding of key aspects fostering the exposure of dairy cows to these pathogens. In the present study, multiple multinomial logistic regression models were fitted via neural networks to describe the differences among farms where cattle were not exposed to either F. hepatica or O. ostertagi, to one parasite, or to both, respectively. Farm-specific production and management characteristics were used as covariates to portray these differences. This elucidated inherent farm characteristics associated with parasite exposure. In both studied regions, pasture access for cows, farm-level milk yield, and lameness prevalence were identified as relevant factors. In region 'South', adherence to organic farming principles was a further covariate of importance. In region 'North', the prevalence of cows with a low body condition score, herd size, hock lesion prevalence, farm-level somatic cell count, and study year appeared to be of relevance. The present study broadens our understanding of the complex epidemiological scenarios that could predict differential farm-level parasite status. The analyses have revealed the importance of awareness of dissimilarities between farms in regard to the differential exposure to F. hepatica and O. ostertagi. This provides solid evidence that dynamics and relevant factors differ depending on whether or not cows are exposed to F. hepatica, O. ostertagi, or to both.

Detection of Toxoplasma gondii DNA in milk of dairy cows from southern Brazil.

Consumption of unpasteurized cow's milk may be a transmission route for some pathogenic microorganisms, but there is little information about the risk of Toxoplasma gondii infection. Blood and milk samples were collected in a paired and random fashion from 106 dairy cows and bulk-tank milk samples were also collected from each of the six farms, in southern Brazil. Serum anti-T.gondii antibodies (IgG) were detected by an indirect fluorescent antibody test (IFAT) with a cutoff point of 1:64. Nested PCR targeting the ITS1 was performed on milk samples to detect the Sarcocystidae family, confirmed to be T.gondii by Sanger sequencing. The occurrence of anti-T.gondii antibodies in the herds was 14.1%, (15/106) with seropositive cows in all herds. Antibody titers in positive samples ranged from 64 to 128. T.gondii DNA was detected in 2.8% (03/106) of the milk samples. The ITS1 sequences generated in this study were ON809793 - ON809794 and the sequencing revealed 98-100% identity with T. gondii DNA sequences deposited in GenBank. All cows PCR positive for T.gondii in milk were negative for IgG antibodies in serum, suggesting that naturally infected cows may shed T. gondii in milk in the acute phase of infection. The results of this study demonstrate that T. gondii DNA may be detected in raw cow's milk, so the potential risks of lactogenic infection should be considered. The presence of T. gondii DNA in milk does not confirm that the protozoa are viable and infective, and further investigations into the role of cow's milk in the epidemiology of toxoplasmosis are needed.

Genetic and seasonal variations of Trypanosoma theileri and the association of Trypanosoma theileri infection with dairy cattle productivity in Northern Japan.

Trypanosoma theileri is considered a non- or low-pathogenic trypanosome that generally causes latent infection in apparently healthy cattle; however, T. theileri propagates in the bloodstream and may cause clinical disease in pregnant animals or co-infection with bovine leukemia virus or Theileria orientalis. In the current study, a monthly survey of T. theileri infection over one year was carried out in a research dairy farm in Hokkaido, Japan to determine the 1) seasonal variations in the prevalence, 2) genetic characterization of T. theileri, and 3) associations of milk and blood parameters in dairy cattle with T. theileri infection, including data of metabolic profile tests and dairy herd performance tests, using linear mixed models. We found that 1) the prevalence of T. theileri infection was significantly higher in summer and winter than in other seasons; 2) T. theileri possibly showed genetic diversity in Eastern Hokkaido; and 3) T. theileri infection was associated with significantly lower levels of blood urea nitrogen, milk protein, and solids-not-fat, which are caused by a low rumen fermentation level. This is the first study to report the negative impact of T. theileri infection in dairy cattle, and our study indicates that control of T. theileri infection can improve the productivity of dairy cattle.

Herd-level contamination of Neospora caninum, Toxoplasma gondii and Brucella in milk of Iranian dairy farms.

The bulk milk examination is a reliable screening tool for monitoring the quality of milk in the farms. The infection to Neospora caninum, Toxoplasma gondii and Brucella sp. Was evaluated in bulk milk samples of dairy farms in Hamedan province, West part of Iran. All the dairy farms (n = 149) were examined for N. caninum, T. gondii and Brucella infections using milk ring test (MRT), microbiology, serology (Enzyme-linked Immunosorbent Assay), and molecular techniques. Based on molecular methods, Brucella-infection was negative in all farms; while, 55 %, 5.4 % and 2.7 % of samples were positive for N. caninum, T. gondii and mix infection, respectively. The highest Neospora-infection was detected in the farms with history of abortion in fall and winter. There was significant association between Neospora-infection and the presence of dogs and rodents in the farms, herd size, and age of the animals. Also, a significant association was seen between Toxoplasma-infection and the presence of cats and rodents in the farms, as well as age of the animals. Average total bacterial count (TBC) was calculated 1.14 × 106±1.1 × 106. The highest TBC was in the farms from Central locations of studied area (5.7 × 106±2.24 × 106), farms with more than 120 animals (7.9 × 106±2.8 × 106), and farms with ≥50-months age (1.74 × 106±6.3 × 105) in spring and summer (6.9 × 106±3.7 × 106). The number of somatic cells was estimated between 1 × 104 and 2 × 106 (Average = 4.2 × 105±3.39 × 105). The current study was a comprehensive evaluation of Neospora, Toxoplasma and Brucella infections in milk samples of Iranian dairy farms for the first time. Neospora-infection is responsible for economic losses in the region. Health education and milk pasteurization are so helpful for inhibiting the milk borne diseases. To reduce the risk factors, predict and design the appropriate schemes like redundant of heterogeneous animals are recommended.

Public Health Significance of Toxoplasma gondii Infections in Cattle: 2009-2020.

Toxoplasma gondii infections are common in humans and animals worldwide. The ingestion of food or water contaminated with oocysts excreted by infected cats or ingesting uncooked or undercooked meat containing tissue cysts of T. gondii are the 2 major modes of transmission of T. gondii. Viable T. gondii is more prevalent in pork and lamb than in beef. In the past decade, there have been many articles on the high seroprevalence in cattle, particularly from China. There is a report of an outbreak of acute toxoplasmosis in humans suspected to be linked to the ingestion of Artisan fresh cheese from cow's milk. There are conflicting reports concerning the rate of congenital transmission of T. gondii in cattle, especially from Brazil. In a report from Brazil, viable T. gondii was isolated from the blood of 1 of 60 pregnant cows slaughtered at an abattoir and from 1 fetus. The role of beef in the epidemiology of T. gondii infections is still not clear. Here, we review prevalence, persistence of infection, clinical disease, epidemiology, and public health risks of T. gondii infections in cattle from beef and cow's milk worldwide for the past decade.

Prototheca from bovine milk and associated minimal algaecide concentration of chlorhexidine and povidone-iodine in Taiwan.

OBJECTIVE: Protothecal mastitis has emerged and affected the dairy industries globally in last decades; however, little is known about the occurrence of Prototheca in Taiwan. The present study identified the Prototheca isolated from bovine milk and revealed the algaecide efficacy of 2 disinfectants. MATERIAL AND METHODS: From 2011 to 2014, a total of 176 Prototheca isolates were obtained from 10 625 milk samples collected from dairy cows in 24 Taiwanese dairy farms. Of these, 149 isolates were identified by biochemical tests and genotype differentiation with a specific PCR protocol. Minimal algaecide concentration (MAC) of chlorhexidine and povidone-iodine were obtained from 16 isolates by microdilution method. RESULTS: Almost all (n = 147) isolates were P. zopfii, and remaining 2 isolates were P. wickerhamii and unidentified, respectively. Based on molecular genotyping, all P. zopfii isolates belonged to genotype 2. The MAC of 2 commonly used disinfectants, chlorhexidine and povidone-iodine, against 16 P. zopfii isolates was 1.56-3.13 and 48.83-390.63 µg/ml, respectively. CONCLUSION: This report revealed for the first time the prevalence and the predominant P. zopfii genotype 2 of bovine protothecal infection in Taiwan. In vitro algaecide efficacy indicated that both tested disinfectants were effective against P. zopfii.

Risk factors for Toxoplasma gondii seropositivity in the Old Order Amish.

Toxoplasma gondii (T. gondii) is an important human disease-causing parasite. In the USA, T. gondii infects >10% of the population, accrues economic losses of US$3.6 billion/year, and ranks as the second leading culprit of foodborne illness-related fatalities. We assessed toxoplasmosis risk among the Old Order Amish, a mostly homogenous population with a high prevalence of T. gondii seropositivity, using a questionnaire focusing on food consumption/preparation behaviours and environmental risk factors. Analyses were conducted using multiple logistic regression. Consuming raw meat, rare meat, or unpasteurised cow or goat milk products was associated with increased odds of seropositivity (unadjusted Odds Ratios: 2.192, 1.613, and 1.718 , respectively). In separate models by sex, consuming raw meat, or consuming unpasteurised cow or goat milk products, was associated with increased odds of seropositivity among women; washing hands after touching meat with decreased odds of seropositivity among women (adjusted OR (AOR): 0.462); and cleaning cat litterbox with increased odds of seropositivity among men (AOR: 5.241). This is the first study to assess associations between behavioural and environmental risk factors and T. gondii seropositivity in a US population with high seroprevalence for T. gondii. Our study emphasises the importance of proper food safety behaviours to avoid the risk of infection.

Arcobacter butzleri, un agente zoonótico en leche bovina / Arcobacter butzleri, a zoonotic agent in bovine milk / Arcobacter butzleri, um agente zoonótico em leite Bovino

Para la búsqueda de especies de Arcobacter fueron estudiadas 50 muestras de leche bovina obtenidas de los centros de acopio de la empresa láctea Ecolac, de las provincias de Loja y Zamora Chinchipe, Ecuador, y se aisló un total de ocho cepas de Arcobacter butzleri (16%). Esta frecuencia de aislamiento concuerda y está dentro de los rangos descriptos en la literatura. Todas las cepas aisladas fueron sensibles a gentamicina. Se encontró alta frecuencia de resistencia a tetraciclina (6/8 cepas) y a ciprofloxacina (4/8 cepas). Se verificó la ocurrencia de multirresistencia en tres de las ocho cepas aisladas.

A total of 50 samples of bovine milk obtained from bulk tanks milk of the collection centers belonging to the company ECOLAC, of the provinces of Loja and Zamora Chinchipe, Ecuador, were studied for Arcobacter species diagnosis, being isolated 8 strains of Arcobacter butzleri (16%). This frequency of isolation agrees and falls within the ranges described in the literature. All the isolated strains were susceptible to gentamicin. High resistance levels to tetracycline and ciprofloxacin were found with 6/8 and 4/8 resistant strains respectively. Multi-resistance was found in three of the eight isolated strains.

Foram estudadas, para a pesquisa de espécies de Arcobacter, 50 amostras de leite bovino, obtidas dos centros de coleta da empresa de laticínios ECOLAC, das províncias de Loja e Zamora Chinchipe, Equador, sendo isoladas em total 8 cepas de Arcobacter butzleri (16%). Esta frequência de isolamento concorda e está dentro dos níveis descritos na literatura. Todas as cepas isoladas foram sensíveis à gentamicina. Foi encontrada alta frequência de resistência à tetraciclina (6/8 cepas) e à ciprofloxacina (4/8 cepas), sendo verificada a ocorrência de multirresistência em três das oito cepas isoladas.

Molecular and microscopic detection of natural and experimental infections of Toxocara vitulorum in bovine milk.

Toxocara vitulorum is an Ascarid nematode infecting the small intestine of buffalo and cattle particularly neonate calves, with the postnatal route through milk is the main infection source. However, little is known about shedding rates and the optimum detection methods of T. vitulorum larvae in the milk of the infected bovine hosts. In this study, we aimed to evaluate the use of two methods, microscopy and PCR, and their detection limits both under the experimental and natural infection situations. In doing this, T. vitulorum eggs extracted from naturally occurring adult female worms were successfully subjected to experimental embryonation, and larvae were implemented in experimental infection of milk in ascending infection doses of 0, 1, 5, 10, 20, 50 larvae/2-ml milk samples. With the except of negative control, microscopy-based examination detected larvae in all samples, albeit with means, ranges, and the total number of larvae were detected in exponential rates relative to larvae densities in milk samples. PCR technique corresponded well to microscopy in detecting genomic DNA of T. vitulorum larvae in all milk samples down to a single larva/sample. On the other hand, and by applying the same methodology approach on 50 naturally-occurring bovine colostrum/milk samples, 13 (26%) and 20 (40%) samples were tested positive for T. vitulorum infection by microscopy and the PCR-based detection, respectively. Of these, 11 out of 26 buffalo samples (42.30%) and 2 out of 24 cow samples (8.33%) were tested positive by microscopy, while 16 (61.54%) and 3 (12.50%) of buffalo and cow samples were tested positive by PCR, respectively. By applying the Agreement Coefficient, substantial agreement (0.77) between molecular and microscopy detection was detected from all tested samples. In conclusion, larvae of T. vitulorum were unequivocally detected by microscopy and molecular methods in milk samples both under the experimental and natural field situations. Nevertheless, slightly higher rates by PCR than microscopy were obtained when detecting naturally-infected milk samples. To the best of our knowledge, this is the first in situ detection of larvae of T. vitulorum in the milk of the naturally infected animals.

A review of outbreaks of cryptosporidiosis due to unpasteurized milk.

PURPOSE: This review analysed outbreaks of human cryptosporidiosis due to raw milk. The objective of our study was to highlight and identify underestimated and underreported aspects of transmission of the parasite as well as the added value of genotyping Cryptosporidium isolates. METHODS: We conducted a descriptive literature review using the digital archives Pubmed and Embase. All original papers and case reports referring to outbreaks of Cryptosporidium due to unpasteurized milk were reviewed. The cross-references from these publications were also included. RESULTS: Outbreaks have been described in the USA, Australia, and the UK. Laboratory evidence of Cryptosporidium from milk specimens was lacking in the majority of the investigations. However, in most recent reports molecular tests on stool specimens along with epidemiological data supported that the infection was acquired through the consumption of unpasteurized milk. As the incubation period for Cryptosporidium is relatively long (days to weeks) compared with many other foodborne pathogens (hours to days), these reports often lack microbiological confirmation because, by the time the outbreak was identified, the possibly contaminated milk was not available anymore. CONCLUSION: Cryptosporidiosis is generally considered a waterborne intestinal infection, but several reports on foodborne transmission (including through raw milk) have been reported in the literature. Calves are frequently infected with Cryptosporidium spp., which does not multiply in milk. However, Cryptosporidium oocysts can survive if pasteurization fails. Thus, pasteurization is essential to inactivate oocysts. Although cryptosporidiosis cases acquired from raw milk are seldom reported, the risk should not be underestimated and Cryptosporidium should be considered as a potential agent of contamination. Genotyping Cryptosporidium isolates might be a supportive tool to strengthen epidemiologic evidence as well as to estimate the burden of the disease.

Unpasteurised milk consumption as a potential risk factor for toxoplasmosis in females with recurrent pregnancy loss.

In women with a bad obstetric history, certain infections are associated with recurrent foetal loss. One of the common infectious agents is a protozoan parasite, Toxoplasma gondii. The aim of this study was to assess unpasteurised milk consumption as a potential risk factor for toxoplasmosis in females with recurrent pregnancy loss from the province of Khyber Pakhtunkhwa, Pakistan. In this study, we recruited a total of 360 females, comprising a study group of 180 females with previous history of recurrent pregnancy loss and a control group of 180 females with no such history. Blood serum from the participants was analysed for Toxoplasma gondii IgM antibodies by enzyme linked immunosorbent assay. Among the study group, 23 (12.8%) females were serologically positive for IgM antibodies against Toxoplasma gondii, whilst 157 (87.2%) were IgM negative. In the control group, only two (4.8%) females were IgM positive, whilst 178 (95.2%) were IgM negative. Bad pregnancy outcome in the study group and control group was observed to be significantly different (p < .0001). In both of these groups, unpasteurised milk consumption was found as a major risk factor for Toxoplasma gondii infection. A routine serological investigation should be carried out in pregnant women to rule out toxoplasmosis and reduce the risk of recurrent pregnancy loss as well as congenital toxoplasmosis in newborns.Impact statementWhat is already known on this subject? Seropositivity for Toxoplasma gondii antibodies ranges from 7% to 51% in different regions of the world. The prevalence rate varies because of differences in climate, culture, food habits, behaviour, personal hygiene and cooking habits of different societies and ethnic groups. Various risk factors have been identified that contribute to a high prevalence rate of the disease, including consumption of raw or poorly cooked meat, physical contact with cats or cat litter, consumption of unwashed raw vegetables and fruits, drinking of contaminated water and milk. We presumed that consuming unpasteurised milk could be a potential risk factor for developing toxoplasmosis in pregnant women.What the results of this study add? This study demonstrates high seroprevalence of Toxoplasma gondii antibodies in females of child bearing age that have consumed unpasteurised milk and is a potential risk factor for developing toxoplasmosis.What the implications are of these findings for clinical practice and/or further research? Our findings suggest that primary preventive measures (personal hygiene, frequent hand washing and consuming pasteurised milk) should be taken by health surveillance authorities to focus on families, especially pregnant women, to educate them about personal hygiene, contact with cattle or using their milk and milk products. The latter is especially important to aware them about the hazards of consuming unpasteurised and contaminated milk.

Seroprevalence of Toxoplasma gondii among pregnant women attending antenatal clinics at Hawassa University comprehensive specialized and Yirgalem General Hospitals, in Southern Ethiopia.

BACKGROUND: Toxoplasmosis is caused by infection with the protozoan parasite Toxoplasma gondii. It is acquired by consumption of raw or undercooked meat containing tissue cyst, food or water contaminated with oocyst and congenital infection through the placenta leading to serious congenital abnormalities in the fetus like miscarriage, stillbirth, intrauterine death and neurologic defects. Therefore; this study was aimed to determine the prevalence of toxoplasmosis infection and its possible risk factors associated with pregnant women attending antenatal clinics in Hawassa and Yiregalem Hospitals, Southern Ethiopia. METHODS: A hospital-based cross-sectional study was conducted from December 2016 to May 2017. The study was done in antenatal care clinics of Hawassa and Yiregalem Hospitals in Southern, Ethiopia. Five hundred pregnant women were interviewed with a pretested structured questionnaire to collect risk factors and socio-demographic data. Blood samples were collected and serum was separated and tested for anti- Toxoplasma gondii antibodies using ELISA (Enzyme-linked Immunosorbent Assay). Data were analyzed using SPSS version 20 statistical software. The risk factors were tested for significance using Bivariate and multivariate analysis. P-value < 0.05 was considered statistically significant. RESULTS: The weighted prevalence of this study was 81.8% for the anti- Toxoplasma gondii antibody. Almost all participants (99.6%) had no information about the disease. A significant association was observed between seroprevalence and contact with domestic cats (OR = 1.206, 95% CI (1.627-2.206, P = 0.043), consumption of raw meat (OR = 0.848, 95% CI: 1.517-2.941, P = 0.019) and unpasteurized milk (OR = 0.871, 95% CI 1.531-2.221, P = 0.032). A significant association was not observed between seroprevalence and age, history of abortion, and blood transfusion. CONCLUSIONS: The findings of this study demonstrated a relatively higher prevalence of seropositivity than studies reported from other countries. Existence of domestic cats at home, consumption of undercooked meat and unpasteurized milk were identified as risk factors for T. gondii infection. Therefore, a health education program to increase the mother's knowledge about toxoplasmosis towards avoiding eating undercooked meat, contact with cats and consumption of unpasteurized milk during pregnancy is recommended. Furthermore, our results suggested that the implementation of newborn screening and follow-up testing can lead to reducing of toxoplasmosis associated complications.

Toxoplasma gondii in naturally infected goats: Monitoring of specific IgG levels in serum and milk during lactation and parasitic DNA detection in milk.

The zoonotic protozoa Toxoplasma gondii is one of the major abortive pathogens in small ruminants. Nevertheless, data on T. gondii infection in goats during lactation and on the presence of T. gondii in goat milk are lacking. A longitudinal study was planned in a T. gondii naturally infected dairy goat farm with the aim of (i) evaluating the variation of anti-T. gondii antibodies in blood and milk during the lactation; (ii) identifying the optimal phase during lactation for T. gondii monitoring; (iii) detecting the presence of T. gondii DNA in the milk. From March to July 2017, 30 goats in a farm were fortnightly visited seven times and sampled for blood and, when in lactation, for milk. Individual data regarding age, reproductive disorders, and the day of lactation were recorded. For the detection of anti-T. gondii antibodies in blood and milk a commercial ELISA kit was used. Milk samples (n = 63) of selected nine seropositive animals were also molecularly analysed to amplify a sequence within the ITS1 region of T. gondii. The seroprevalence of T. gondii infection was 63.3% (19/30); a high agreement was obtained between serum and milk results (Spearman's coefficient = 0.793 and Kendall's tau = 0.624), particularly between the 15th and the 60th day of lactation. In the statistical analysis, performed with generalized linear mixed models (GLMMs), the variable "phase of lactation" was strongly associated to ELISA values obtained in both serum and milk (p-value = 0.0001, F = 5.197, and p-value = 0.016, F = 2.755, respectively). Finally, molecular analyses revealed the presence of parasitic DNA in 20.6% (13/63) of milk samples, with a discontinuous parasite excretion; statistical analyses did not reveal any association among the parasite excretion and the considered variables. Milk could be considered as a valid alternative to blood for monitoring T. gondii infection in goat herds. Moreover, the detection of T. gondii DNA in milk enhanced the possibility for raw goat's milk consumption to be considered as a risk to public health.

Detection and genotypic characterization of Toxoplasma gondii DNA within the milk of Mongolian livestock.

Toxoplasma gondii is a global, zoonotic parasite capable of infecting any warm-blooded host. Toxoplasmosis can cause a variety of illnesses including abortions and congenital defects in humans, sheep, and goats. Congenital toxoplasmosis is considered to have the highest global disease burden of any foodborne illness in humans. This study examined the potential role of milk as a route of T. gondii transmission between livestock and humans within Mongolian herders, a little-studied population which relies heavily on animals. Milk of Mongolian sheep, goats and Bactrian camels was tested for the presence of T. gondii DNA, and a survey was conducted to ascertain what behavioral and environmental factors were present that might potentiate T. gondii infection within these Mongolian communities. T. gondii DNA was detected in samples from one sheep and five camels. Sequence analysis of DNA from camel milk revealed that two were from potentially virulent T. gondii genotypes. This has implications for public health in the region, as milk is an extremely important source of nutrition and our survey results imply that some people believe consumption of raw camel milk carries health benefits. This is the first report of T. gondii DNA in Bactrian camel milk as well as the first genotypic characterization of T. gondii within Mongolia.

Short communication: Occurrence and persistence of Prototheca zopfii in dairy herds of Korea.

Bovine mastitis caused by Prototheca has been reported globally, and its incidence is increasing in dairy herds. The present study aimed to investigate the occurrence of Prototheca and persistence of Prototheca zopfii strains in Korean dairy herds. A total of 187 (7.5%) P. zopfii strains were isolated from 2,508 quarter milk samples collected from 50 dairy farms throughout Korea from 2015 to 2017. Prototheca zopfii was isolated from one farm among the 50 farms over the 3-yr period. The P. zopfii isolates belonged to genotype 2. Overall, Prototheca-positive quarter milk samples showed high somatic cell counts with an average value of log 6.48 ± 6.54 cells/mL. Prototheca zopfii was found to be persistent in an infected farm over a 2-yr period. To the best of our knowledge, this is the first report on the presence and persistence of protothecal mastitis caused by P. zopfii genotype 2 in a Korean dairy herd. This disease leads to a significant increase in somatic cell counts in milk, which persists for more than 1 yr in the affected cow udder. These results suggest that P. zopfii could pose a serious risk to dairy herds. Thus, strict surveillance for protothecal mastitis is urgently needed and sanitary conditions regarding the environment and milk collection are essential because of the lack of effective treatment options.

Seroprevalence of Toxoplasma gondii infection in children with central nervous system disorders in Mansoura, Egypt: a case-control study.

Background: Toxoplasma gondii is a global infection with a crucial role in the development of neurological diseases. Data concerning the association between T. gondii and neurological illnesses in Egyptian children is scarce. Methods: A case-control study was conducted on 60 patients divided into children suffering from central nervous system manifestations without apparent chromosomal anomalies (n=30) and children with Down syndrome (n=30) recruited from Mansoura University Children's Hospital, Mansoura, Egypt. A total of 30 healthy children were included as controls. Demographics and clinical data were collected from all cases and Toxoplasma immunoglobulin (Ig) M and G antibodies were assessed by enzyme-linked immunosorbent assay. Results: Anti-T. gondii IgG was the most frequent antibody detected and the highest seropositivity rates were ranked for the neurologically disabled non-syndromic children, followed by Down syndrome, compared with controls (p≤0.001). Statistically significant (p=0.05) associations were found between Toxoplasma IgG seropositivity and hydrocephalus and between Toxoplasma IgM and a history of contact with farm animals, soil and cats in children with Down syndrome. Conclusions: The association between Toxoplasma infection and neurological disorders in children should be kept in mind by paediatricians and assessment of T. gondii antibodies in early childhood is needed for timely management of afflicted patients.

Herd-level risk factors associated with Brucella sero-positivity in cattle, and perception and behaviours on the disease control among agro-pastoralists in Tanzania.

Brucellosis is endemic in Tanzania, particularly in agro-pastoral areas. This study investigated the herd-level sero-prevalence and risk factors for Brucella sero-positivity in cattle, and perception and behaviours associated with brucellosis control among agro-pastoralists in Morogoro Region, Tanzania. A cross-sectional study involving herd milk diagnosis by indirect enzyme-linked immunosorbent assay and questionnaire survey was conducted in 124 farms. Questions included potential risk factors, knowledge of brucellosis, willingness-to-pay for cattle vaccination, and item count technique (ICT) for selling behaviour of cows that experienced abortion. Risk factor analysis for Brucella sero-positivity in cattle and analysis of factors associated with willingness-to-pay were conducted using classical tests and generalised linear models. Most farmers had little knowledge about brucellosis (disease name: 13.7%, symptoms: 3.2%, transmission from cattle to human: 2.4%, and Brucella vaccine: 2.4%). The proportion of Brucella sero-positive herd was 44.4% (55/124, 95%CI: 35.5-53.5). No risk factors for Brucella sero-positivity were identified; however, using a veterinary service was identified as a preventive factor (OR = 0.39, 95%CI: 0.18-0.84, p =  0.02). For scenarios of vaccinating all cattle and only calves, 59.7% and 89.5% of farmers were willing to pay for vaccination, respectively. Being a Maasai tribe member was a hesitating factor for vaccinating all cattle (OR = 0.39, 95%CI: 0.19-0.83, p =  0.01) and using a veterinary service was an encouraging factor for vaccinating calves (OR = 4.0, 95%CI: 1.2-13.0, p =  0.02). The ICT found that 45.1% of farmers sold cows that experienced abortion. This estimate was not statistically different from that obtained by direct questioning (34.1%, SE = 7.5%, binomial p value = 0.27, factor score = 1.32), suggesting that farmers did not hesitate to sell such cows. The Maasai conducted more risky behaviours for human infection such as drinking raw milk (p = 0.06) or blood (p <  0.01) and helping delivery with bare hands (p = 0.03) than other tribes. Community-based brucellosis control programmes with calf vaccination may be feasible in the study areas. A One Health approach including the promotion of health education and expansion of veterinary services is crucial for disease control.

Toxoplasma gondii Antibodies in Bulk Tank Milk Samples of Caprine Dairy Herds.

A major public health issue, Toxoplasma gondii infection can affect humans mainly via the consumption of animal products from certain species, including small ruminants. Therefore, a regular monitoring of the infection in ovine and caprine populations is advisable for the control of human and animal toxoplasmosis. Antibody detection in individual and bulk tank milk (BTM) may represent a valid alternative to serological analysis, in that its collection is easy and does not affect animal welfare. Many serological tools for milk analysis have already been validated for several parasites, including Apicomplexa. Thus, the aim of the present study was to obtain epidemiological data on T. gondii infection through the detection of antibodies in BTM of dairy goat herds from an important area for caprine dairy production (northern Italy). The performance of a commercial ELISA was first evaluated for analysis of caprine milk samples, using a panel of serum-milk pairs of goats naturally infected by T. gondii. The analysis of BTM confirmed the presence of anti- T. gondii antibodies in 59% of the samples. Toxoplasma gondii antibody positivity was more frequently found in goats reared on farms under extensive (64.9%) or semi-intensive systems (68.7%) in comparison with intensive farms (51.1%). Analysis of milk was a valid alternative to serological tests, being easily applied in large-scale epidemiological surveys and for continuous monitoring of T. gondii infection.