Microbial safety status of Serro artisanal cheese produced in Brazil.

Considering the growing consumption of artisanal foods worldwide, we aimed to evaluate the microbial safety of Serro artisanal cheese (SAC), produced in Minas Gerais State, Brazil. This cheese is produced with raw milk using 1 of 2 natural starter cultures: "pingo" and "rala." A total of 53 SAC samples (pingo = 8; rala = 45) were obtained from different farmers and subjected to conventional and molecular assays to detect and enumerate Listeria monocytogenes, Salmonella spp., coagulase-positive staphylococci (CPS), diarrheagenic Escherichia coli, Mycobacterium tuberculosis, and Brucella abortus. The SAC samples were also subjected to an ELISA to detect classical staphylococcal enterotoxins (CSE: SEA, SEB, SEC, SED, SEE) and to PCR assays to detect staphylococcal enterotoxin-related genes (sea, seb, sec, sed, see). Coagulase-positive staphylococci isolates were obtained and tested by the same assays to detect their potential in CSE production and presence of CSE-related genes. None of the SAC samples showed any of the screened food-borne pathogens and zoonotic agents, and none showed the presence of CSE by phenotypic and genotypic approaches. Despite the absence of microbial hazards, mean counts of CPS in SAC samples were 5.2 log cfu/g (pingo starter) and 4.6 log cfu/g (rala starter), indicating poor hygiene practices during production. None of the tested CPS isolates (n = 116) produced CSE or presented CSE-related genes. Despite the relative microbial safety, hygienic conditions during SAC production must be improved to meet official guidelines established in Brazil.

Detection and survival of Toxoplasma gondii in milk and cheese from experimentally infected goats.

The consumption of unpasteurized goat cheese and goat's milk has been suggested as a risk factor for toxoplasmosis in humans. In the present study, detection and survival of Toxoplasma gondii in milk and cheese was studied by bioassay in mice (milk) and in cats (cheese). Eight goats were inoculated orally with 300 to 10,000 oocysts of T. gondii strain TgGoatUS26. Milk samples were collected daily up to 30 days postinoculation and bioassayed in mice and cats. For mouse bioassay, 50 ml of milk samples were centrifuged, and the sediment was inoculated subcutaneously into mice. Mice were tested for T. gondii infection by seroconversion and by the demonstration of parasites. By mouse bioassay, T. gondii was detected in milk from all eight goats. The T. gondii excretion in milk was intermittent. For cat bioassay, 400 ml (100 ml or more from each goat) of milk from four goats from 6 to 27 days postinoculation were pooled daily, and cheese was made using rennin. Ten grams of cheese was fed daily to four cats, and cat feces were examined for oocyst shedding. One cat fed cheese shed oocysts 7 to 11 days after consuming cheese. Attempts were made to detect T. gondii DNA in milk of four goats; T. gondii was detected by PCR more consistently, but there was no correlation between detection of viable T. gondii by bioassay in mice and T. gondii DNA by PCR. Results indicate that T. gondii can be excreted in goat's milk and can survive in fresh cheese made by cold-enzyme treatment. To prevent transmission to humans or animals, milk should not be consumed raw. Raw fresh goat cheese made by cold-enzyme treatment of unpasteurized milk also should not be consumed.

Surveillance of feral swine for Trichinella spp. and Toxoplasma gondii in the USA and host-related factors associated with infection.

Trichinella spp. and Toxoplasma gondii are important zoonotic parasites that infect warm blooded animals and humans worldwide. Among domesticated food animals, pigs are the main host for Trichinella spiralis. Pigs, chickens, sheep, and goats are known to be infected with T. gondii at varying rates, depending on husbandry. Infections in wildlife with these parasites are generally higher than in domesticated species. Feral swine act as reservoirs of infection in the sylvatic ecosystem for Trichinella spp. and T. gondii, acting as sources of infection for peridomestic carnivores whose home ranges overlap with domestic pigs. Feral swine can have direct contact with non-biosecure domestic pigs, presenting opportunity for direct disease transmission through cannibalistic behavior. Determination of the prevalence of Trichinella spp. and T. gondii infection in feral swine is needed to understand the risk of transmission of these parasites to domestic pigs. A cross-sectional serological survey was conducted between 2006 and 2010 to estimate the antibody prevalence of Trichinella spp. and T. gondii and risk factors associated with infection in feral swine in the USA. Serum samples were tested from 3247 feral pigs from 32 states; results are reported from 26 states. Maximum entropy ecological niche modeling and spatial scan statistic were utilized to predict the geographic range and to examine clusters of infection of Trichinella spp. and T. gondii in feral pigs. The seroprevalence of antibodies to Trichinella spp. and T. gondii was 3.0% and 17.7%, respectively. Species distribution modeling indicated that the most probable distribution areas for both parasites was similar, concentrated primarily in the South and the Midwest regions of the USA. A follow up survey conducted during 2012-2013 revealed that 2.9% of 984 sampled feral swine were seropositive for Trichinella spp., and 28.4% were seropositive for T. gondii. Three hundred and thirty (330) tongues were collected from the 984 sampled animals during 2012-2013; 1.81% were tissue positive for T. spiralis muscle larvae; no other genotypes were found. The potential exists for introduction of these pathogens into domestic herds of non-biosecure domestic pigs as a result of increasing overlap of the range of feral pigs with non-biosecure domestic pigs production facilities in the USA.

Epidemiology of toxoplasmosis in white tailed deer (Odocoileus virginianus): occurrence, congenital transmission, correlates of infection, isolation, and genetic characterization of Toxoplasma gondii.

The prevalence of Toxoplasma gondii in white tailed deer (WTD) in the USA is high but little is known of the epidemiology of toxoplasmosis in this host. In the present study, we compared T. gondii seroprevalence from 749 WTD collected in 2012 and 2013 from a Metropolitan Park in Ohio and 487 WTD deer shot in Minnesota during 2008, 2009, and 2010. Serum samples were tested for antibodies to T. gondii by the modified agglutination test (cut-off titer, 25). Additionally myocardial samples from 123 seropositive WTD from Ohio were digested in pepsin and the digests were bioassayed for the isolation of T. gondii. Furthermore, to estimate transplacental rate of transmission, brains from 155 fetuses (included twins) from 148 deer from Minnesota were bioassayed in mice for the isolation of viable T. gondii. Seroprevalence of T. gondii varied with the year of collection, geography, and the age of deer. Of the Ohio deer sampled in 2012 and 2013 seroprevalences for the two years were similar (73.4% and 75.7%, respectively); remarkably 150 (66.1%) of 227 deer of <1 year of age were seropositive. Of the Minnesota deer, seroprevalence was lowest for the year 2008 (14.8%, 26/175) versus 2009 (27.7%, 59/213), and 2010 (25.2%, 25/99), thought to be related to environmental temperatures. Viable T. gondii was isolated in mice from the myocardium of four WTD from Ohio, and brain of one WTD fetus from Minnesota. Tachyzoites from infected mouse tissues were further propagated in cell culture. The DNA isolated from culture-derived tachyzoites of these five T. gondii isolates was characterized using 11 PCR-RFLP markers (SAG1, 5'- and 3'-SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico). Four genotypes were found, including ToxoDB genotype no. 1 (Type II), no. 2 (Type III), no. 3 (Type II variant) and no. 146. Results indicate fluctuating seroprevalence, probably related to weather and warrant further epidemiological studies.

Isolation of viable Neospora caninum from brains of wild gray wolves (Canis lupus).

Neospora caninum is a common cause of abortion in cattle worldwide. Canids, including the dog and the dingo (Canis familiaris), the coyote (Canis latrans), and the gray wolf (Canis lupus) are its definitive hosts that can excrete environmentally resistant oocysts in the environment, but also can act as intermediate hosts, harboring tissue stages of the parasite. In an attempt to isolate viable N. caninum from tissues of naturally infected wolves, brain and heart tissue from 109 wolves from Minnesota were bioassayed in mice. Viable N. caninum (NcWolfMn1, NcWolfMn2) was isolated from the brains of two wolves by bioassays in interferon gamma gene knockout mice. DNA obtained from culture-derived N. caninum tachyzoites of the two isolates were analyzed by N. caninum-specific Nc5 polymerase chain reaction and confirmed diagnosis. This is the first report of isolation of N. caninum from tissues of any wild canid host.

Genotyping Toxoplasma gondii from wildlife in Pennsylvania and identification of natural recombinants virulent to mice.

Recent studies indicated the predominance of Toxoplasma gondii haplogroup 12 in wildlife in the USA. However, still little is known of the genetic diversity of this parasite circulating in wildlife. In the present study, we tested coyotes (Canis latrans), red foxes (Vulpes vulpes), white-tailed deer (Odocoileus virginianus), and geese (Branta canadensis) from the state of Pennsylvania for T. gondii infection. Antibodies to T. gondii were found in 160 of 367 animals, including 92 (34.5%) of 266 coyotes, 49 (62.0%) of 79 white-tailed deer, 17 (85.0%) of 20 red fox, and two of two Canada geese tested by the modified agglutination test (cut off titer 1:25). Tissues from 105 seropositive animals were bioassayed in mice, and viable T. gondii was isolated from 29 animals, including 10 of 53 coyotes, 11 of 16 foxes, 7 of 49 deer, and one of one goose. DNA isolated from culture-derived tachyzoites of these isolates was characterized initially using multilocus PCR-RFLP markers. Nine genotypes were revealed, including ToxoDB PCR-RFLP #1 (4 isolates), #2 (2 isolates), #3 (4 isolates), #4 (6 isolates), #5 (4 isolates), #54 (1 isolate), #141 (1 isolate), #143 (1 isolate), and #216 (6 isolates), indicating high genetic diversity of T. gondii in wildlife in Pennsylvania. Pathogenicity of six T. gondii isolates (5 of #216 and #141) was determined in outbred Swiss Webster mice. Three of #216 and the #141 isolates were acute virulent to mice, and the other 2 #216 isolates were intermediate virulent. To determine the extent of genetic variation of these as well as a few recently reported virulent isolates from wildlife in North America, intron sequences were generated. Analysis of intron sequences and PCR-RFLP genotyping results indicated that the #216 isolates are likely derived from recombination of the clonal type I and III lineages. To determine if T. gondii virulence can be predicted by typing, we genotyped a collection of strains using PCR-RFLP markers for polymorphic genes ROP5, ROP16, ROP18 and GRA15, which are known to interact with host immune response. The results showed that there is an association of genotypes of ROP5 and ROP18 with mouse-virulence, however, additional gene(s) may also contribute to virulence in distinct T. gondii genotypes.

Isolation and RFLP genotyping of Toxoplasma gondii from the domestic dogs (Canis familiaris) from Grenada, West Indies revealed high genetic variability.

Stray dogs are considered as sentinels in the epidemiology of Toxoplasma gondii because they are carnivores and eat a variety of foods, including garbage. In the present study, tissues and sera of 249 stray dogs (Canis familiaris) from Grenada, West Indies were examined for T. gondii infection. Sera were examined for antibodies to T. gondii by the modified agglutination test (MAT); 89 (35.7%) of 249 were seropositive with titers of 25 in seven dogs, 50 in 22 dogs, 100 in 22 dogs, 200 or higher in 38 dogs. Hearts of 76 seropositive dogs were bioassayed in mice. Viable T. gondii was isolated from 12 dogs; these isolates were designated TgDogGr1 to TgDogGr12. These isolates were further propagated in cell culture and DNA isolated from cell culture derived tachyzoites of 12 isolates was genotyped using 10 PCR-restriction fragment length polymorphism markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). The results revealed six genotypes, including ToxoDB PCR-RFLP #1, #2, #3, #7, #13 and #224, with 1, 6, 1, 2, 1 and 1 isolates, respectively. The result supports previous findings that T. gondii population genetics is highly diverse in Grenada.

Isolation and RFLP genotyping of Toxoplasma gondii from the gray wolf (Canis lupus).

Little is known of the genetic diversity of Toxoplasma gondii circulating in wildlife. In the present study feral gray wolves (Canis lupus) from Minnesota were examined for T. gondii infection. Antibodies to T. gondii were detected in 130 (52.4%) of 248 wolves tested by the modified agglutination test (cut-off titer of 25). Tissues (hearts, brains or both) of 109 wolves were bioassayed in mice for protozoal isolation. Viable T. gondii was isolated from 25 and the isolates were further propagated in cell culture. T. gondii DNA from these isolates was characterized using 10 PCR-RFLP markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico). Four genotypes were detected. Twenty-one isolates were Type 12 (ToxoDB PCR-RFLP genotype #5), 2 were Type II clonal (ToxoDB #1), 1 was Type II variant (ToxoDB #3), and 1 was a new genotype designated as ToxoDB genotype #219.

Occurrence, isolation, and genetic characterization of Toxoplasma gondii from white-tailed deer (Odocoileus virginianus) in New Jersey.

The ingestion of uncooked infected white-tailed deer (WTD) tissues can transmit Toxoplasma gondii infection to humans and mesocarnivores, including cats. In the present study, we tested 264 WTD from New Jersey for T. gondii infection during the 2011-2012 hunting season. Serum samples were tested for antibodies to T. gondii by the modified agglutination test (cutoff titer, 25); 76 (28.7%) of 264 WTD were seropositive. Heart muscle samples from 64 seropositive WTD were digested in pepsin, and the digests were bioassayed for the isolation of T. gondii . Viable T. gondii was isolated in mice from the myocardium of 9 WTD; tachyzoites from infected mouse tissues were further propagated in cell culture. One of the 9 strains was highly virulent for outbred Swiss Webster mice. The DNA isolated from culture-derived tachyzoites of these 9 T. gondii isolates was characterized using 11 PCR-RFLP markers (SAG1, 5'- and 3'-SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). Six genotypes were found, including ToxoDB genotype no. 2 (Type III), no. 3 (Type II variant), no. 4 (Type 12), no. 216, no. 220, and no. 221. The last 2 were new genotypes that were reported for the first time. This is the first report of T. gondii infection in deer from this region of the United States.

Congenital transmission of Neospora caninum in white-tailed deer (Odocoileus virginianus).

Neosporosis is an important cause of bovine abortion worldwide. Many aspects of transmission of Neospora caninum in nature are unknown. The white-tailed deer (Odocoileus virginianus) is considered one of the most important wildlife reservoirs of N. caninum in the USA. During the hunting seasons of 2008, 2009, and 2010, brains of 155 white-tailed deer fetuses were bioassayed in mice for protozoal isolation. Viable N. caninum (NcWTDMn1, NcWTDMn2) was isolated from the brains of two fetuses by bioassays in mice, and subsequent propagation in cell culture. Dams of these two infected fetuses had antibodies to N. caninum by Neospora agglutination test at 1:100 serum dilution. DNA obtained from culture-derived N. caninum tachyzoites of the two isolates with Nc5 PCR confirmed diagnosis. Results prove congenital transmission of N. caninum in the white tailed deer for the first time.

Protein kinase C-mediated ATP stimulation of Na(+)-ATPase activity in LLC-PK1 cells involves a P2Y2 and/or P2Y4 receptor.

ATP-activated P2Y receptors play an important role in renal sodium excretion. The aim of this study was to evaluate the modulation of ATPase-driven sodium reabsorption in the proximal tubule by ATP or adenosine (Ado). LLC-PK1 cells, a model of porcine proximal tubule cells, were used. ATP (10(-6)M) or Ado (10(-6)M) specifically stimulated Na(+)-ATPase activity without any changes in (Na(+)+K(+))-ATPase activity. Our results show that the Ado effect is mediated by its conversion to ATP. Furthermore, it was observed that the effect of ATP was mimicked by UTP, ATPγS and 2-thio-UTP, an agonist of P2Y2 and P2Y4 receptors. In addition, ATP-stimulated Na(+)-ATPase activity involves protein kinase C (PKC). Our results indicate that ATP-induced stimulation of proximal tubule Na(+)-ATPase activity is mediated by a PKC-dependent P2Y2 and/or P2Y4 pathway. These findings provide new perspectives on the role of the effect of P2Y-mediated extracellular ATP on renal sodium handling.

Molecular and biological characterization of first isolates of Hammondia hammondi from cats from Ethiopia.

Toxoplasma gondii oocysts are morphologically and antigenically similar to oocysts of another feline coccidian, Hammondia hammondi. The distinction between H. hammondi and T. gondii is important from an epidemiological perspective because all isolates of T. gondii are potentially pathogenic for humans and animals, whereas H. hammondi is not known to cause clinical disease in any naturally infected intermediate or definitive hosts. In the present report, H. hammondi (designated HhCatEt1 and HhCatEt2) oocysts were found microscopically in the feces of 2 of 36 feral domestic cats (Felis catus) from Addis Ababa, Ethiopia. Oocysts were orally infective to Swiss Webster and gamma interferon gene knockout mice; the inoculated mice developed tissue cysts in their muscles. Laboratory-raised cats fed mouse tissues of infected mice shed H. hammondi oocysts with a prepatent period of 5 days. The DNA extracted from sporulated oocysts reacted with H. hammondi-specific primers, and sequences were deposited in GenBank (accession nos. JX477424, and KC223619). This is the first report of isolation of H. hammondi from cats from the African continent.

Isolation and characterization of new genetic types of Toxoplasma gondii and prevalence of Trichinella murrelli from black bear (Ursus americanus).

Black bears (Ursus americanus) are hosts for two important zoonotic parasites, Toxoplasma gondii and Trichinella spp. and bears are hunted for human consumption in the USA. Little is known of the genetic diversity of T. gondii circulating in wildlife. In the present study, antibodies to T. gondii were found in juice from tongues of 17 (25.7%) of 66 wild black bear from Maryland during the hunting season of 2010 and 2011. Antibodies to T. gondii were assessed by the modified agglutination test. Tongues of 17 seropositive bears were bioassayed in mice and viable T. gondii was isolated from three samples. These three T. gondii isolates (TgBbMd1-3) were further propagated in cell culture and DNA isolated from culture-derived tachyzoites was characterized using 11 PCR-RFLP markers (SAG1, 5'- and 3'-SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico). Results revealed three genotypes. TgBbMd1 is a Type 12 strain (ToxoDB PCR-RFLP genotype #4) and TgBbMd2 is ToxoDB PCR-RFLP genotype #216, and TgBbMd3 is a Type II clonal strain (ToxoDB PCR-RFLP genotype #1). The isolate TgBbMd2 was highly virulent for outbred Swiss Webster mice; all infected mice died of acute toxoplasmosis. Results indicate that mouse virulent strains of T. gondii are circulating in wildlife in the USA. These 66 tongues in addition to tongues collected during hunts in previous years were further investigated for the presence of muscle larvae of Trichinella spp. Tongues from 40 bears in 2005, 41 in 2006, 51 in 2007, 56 in 2008, 68 in 2009, 67 in 2010, and 66 in 2011 were subjected to digestion with pepsin/HCl and microscopic examination. Two bears were infected with Trichinella spp.; one in 2008 and one in 2009. Genotyping of collected muscle larvae revealed that the infecting species in both cases was Trichinella murrelli.

Isolation and genetic characterization of Toxoplasma gondii from mute swan (Cygnus olor) from the USA.

Little is known of the genetic diversity of Toxoplasma gondii circulating in wildlife. In the present study, antibodies to T. gondii were determined in serum samples from 632 mute swans (Cygnus olor) collected from different areas of the USA. Sera were tested by T. gondii modified agglutination test; 54 (8.5%) of 632 samples were seropositive with titers of 25 in 28 sera, 50 in 22 sera, 100 in three samples, and 200 or higher in one swan. Hearts from 14 seropositive swans were bioassayed in mice and viable T. gondii (designated TgSwanUs1-3) were isolated from the hearts of three. These three T. gondii isolates were further propagated in cell culture, and DNA isolated from culture-derived tachyzoites was characterized using 11 PCR-RFLP markers (SAG1, 5'- and 3'-SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico). Results of genotyping revealed that two strains (TgSwanUs1, TgSwanUs2) were Type III (ToxoDB PCR-RFLP genotype #2), and TgSwanUs3 was a new genotype designated here as ToxoDB PCR-RFLP genotype #216. Pathogenicity of oocysts derived from these three strains was determined in Swiss Webster (SW) outbred mice. All mice infected with oocysts and tachyzoites of the atypical isolate (TgSwanUs3) died of acute toxoplasmosis, irrespective of the dose. Oocysts of the remaining two isolates were less pathogenic but differed from each other; 10 oocysts of the TgSwanUs1 killed all inoculated mice whereas 1 million oocysts of the TgSwanUs2 were needed to kill all infected SW mice. Isolation of T. gondii from mute swan indicates that the local waters were contaminated by T. gondii oocysts, and that mouse T. gondii virulent strains are circulating in wildlife. Mute swan is a new host record for T. gondii.

Genetic characterization of viable Toxoplasma gondii isolates from stray dogs from Giza, Egypt.

Stray dogs are considered as sentinels in the epidemiology of Toxoplasma gondii because they are carnivores and eat variety of foods, including garbage. In the present study, tissues and sera of 51 stray dogs (Canis familiaris) from Giza, Egypt were examined for T. gondii infection. Sera were examined for antibodies to T. gondii by the modified agglutination test (MAT); 50 of 51 (98%) were seropositive with titers of 20 in four, 40 in four, 80 in one, 100 in eight, 200 in 17, 400 in 11, 800 or higher in five. Hearts of 43 seropositive dogs were bioassayed in mice. Viable T. gondii was isolated from 22 dogs; these isolates were designated TgDogEg1 to TgDogEg22. DNA isolated from cell culture derived tachyzoites of 22 isolates was genotyped using 10 PCR-restriction fragment length polymorphism markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). The results revealed three genotypes and one mixed infection. The three genotypes are ToxoDB PCR-RFLP #2 (type III, four isolates), #3 (type II variant, 11 isolates), #20 (six isolates), 1 mixed infection. These results revealed the dominance of clonal type II, III and ToxoDB #20 lineages of T. gondii in stray dogs from Giza, Egypt.

Prevalence of Toxoplasma gondii from free-range chickens (Gallus domesticus) from Addis Ababa, Ethiopia.

Prevalence of Toxoplasma gondii in free-range chickens (Gallus domesticus) is a good indicator of the environmental contamination with oocysts because chickens become infected mainly by feeding from ground, feed, or soil contaminated with oocysts. The seroprevalence of T. gondii antibodies in 125 free-range chickens from the Addis Ababa, Ethiopia, was determined. Antibodies to T. gondii were assayed by the modified agglutination test; 48 of 125 (38.4%) chickens were seropositive, with titers of 1:5 in 14, 1:10 in 12, 1:20 in 14, 1: 40 in 3, 1: 80 in 1, 1:160 in 1, 1:320 in 1, and ≥1:640 in 2 chickens. The hearts of 115 chickens were bioassayed for T. gondii infection. Hearts of 72 seronegative (modified agglutination test [MAT] < 1:5) chickens were pooled in 4 groups (20 + 18 + 19 + 15) and fed to 4 T. gondii -free cats; none of these 4 cats shed oocysts in their feces examined 3-21 days after feeding chicken tissues. Hearts of 43 seropositive chickens (MAT ≥ 1:5) were bioassayed individually in mice. Toxoplasma gondii was isolated from only 1 chicken, with a MAT titer of 1:80. This isolate was designated TgCKEt1 and was not pathogenic for outbred mice. Restricted fragment length polymorphism (RFLP) genotyping using 10 loci indicated the TgCKEt1 was ToxoDB polymerase chain reaction-RFLP genotype #1 (Type II clonal). Results of this study indicate very low environmental contamination with T. gondii oocysts around Addis Ababa.

Isolation of viable Toxoplasma gondii from tissues and feces of cats from Addis Ababa, Ethiopia.

Cats are important in the epidemiology of Toxoplasma gondii because they are the only hosts that excrete environmentally resistant oocysts in feces. In the present study, hearts, serum, and feces from 36 feral cats from Addis Ababa area, Ethiopia, were examined for T. gondii infection. Antibodies to T. gondii were determined with the modified agglutination test (MAT, cutoff 1:25); 33 cats were seropositive. Hearts of all 36 cats were homogenized, digested in pepsin, and bioassayed in mice. Feces were examined for T. gondii oocysts by bioassay in mice. Viable T. gondii was isolated from heart of 26 by bioassay in mice and from 25 seropositive and 1 seronegative cats. Toxoplasma gondii was isolated from feces (oocysts) by bioassay in mice. In total, viable T. gondii was isolated from 27 of the 36 cats, and these isolates were designated TgCatEt1 to TgCatEt27. The high prevalence of T. gondii oocysts in feces of 8 (19.4%) of 36 cats is of high epidemiologic significance. This is the first report of isolation of viable T. gondii from any host in Ethiopia.

High prevalence and genotypes of Toxoplasma gondii isolated from organic pigs in northern USA.

The ingestion of undercooked pork infected with Toxoplasma gondii is considered an important source of transmission of this parasite. While T. gondii infection in confinement raised market pigs (market pigs are typically used for fresh, unprocessed pork products) in the USA has decreased significantly over the last 20 years, infection levels in pigs with access to the outdoors can be quite high. An upsurge in consumer demand for 'organically raised', 'humanely raised' and 'free range' pork products has resulted in increasing numbers of hogs being raised in non-confinement systems. To determine T. gondii infection rate in these organic pigs, prevalence of T. gondii in organically raised pigs in two establishments (Farm 1, Farm 2) in Michigan was investigated. Serum and tissue samples from 33 pigs on the farm were available for T. gondii evaluation at slaughter. Serological testing was performed using both ELISA and the modified agglutination test (MAT). Antibodies to T. gondii were detected by both ELISA and MAT in 30 of 33 animals with MAT titers of 1:25 in three, 1:50 in six, 1:100 in seven, 1:200 in 13, and 1:400 in one. Hearts of all 33 pigs were bioassayed for T. gondii in mice; T. gondii was isolated from 17 pigs including one from a seronegative (both ELISA and MAT) pig. Genetic typing of 16 of the 17 T. gondii isolates using the SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico loci revealed clonal Type II from Farm 1 and clonal Type III on Farm 2. These results revealed very high prevalence of T. gondii in organic pigs for the first time in USA, indicating potentially increased health risk of consuming organic swine products.

Chemopreventive potential of the tannase-mediated biotransformation of green tea.

Green tea (Camellia sinensis) is one of the most widely consumed beverages in the world. The cancer chemopreventive qualities of green tea have been well documented. Epigallocatechin gallate (EGCG) is often described as the most potently chemopreventive green tea catechin; however, the low bioavailability of EGCG is a limiting factor for its biological effect. Thus, the aim of this work was to test the chemopreventive potential of green tea extract and EGCG after tannase-mediated hydrolysis. The results showed that the biotransformed compounds retained most of the beneficial properties of the original compounds, and some beneficial properties were improved in the biotransformed compounds. Biotransformation of EGCG decreased its toxicity without affecting its antiproliferative effects. Furthermore, human cells gene expression profiling showed that the biotransformed compounds modulated the expression of several genes related to carcinogenesis. These results demonstrate the benefits of the biotechnological modification of natural food molecules, allowing the improvement of the nutraceutical potential of a beverage as green tea.