Prevalence and molecular identification of Trichinella species isolated from wildlife originating from Limpopo and Mpumalanga provinces of South Africa.

Trichinella species are widely distributed on all continents with the exception of Antarctica, although the full spectrum of Trichinella species found in sub-Saharan African countries, and their hosts, has not been fully documented. This study was conducted to determine the prevalence of Trichinella in wildlife from the Greater Kruger National Park (GKNP) and adjacent areas located in the Limpopo and Mpumalanga provinces of South Africa, and to identify the species and/or genotypes of Trichinella larvae isolated from muscle tissues, using molecular techniques. A review of Trichinella spp. and their wildlife hosts reported during 1964-2011 was also conducted and the results were compared with our current study. Ninety samples representing 15 mammalian, two bird and three reptile species were screened for Trichinella infection during 2012-2016, using artificial digestion. Isolates detected were identified using a multiplex polymerase chain reaction (PCR) amplification of the internal transcriber spacers ITS1 and ITS2, and expansion segment V (ESV) regions of ribosomal DNA, followed by molecular analysis of the sequences. Twenty samples from seven wildlife species were positive for Trichinella spp. larvae, with an overall prevalence of 21.1% (20/90). The prevalence was higher in carnivores (18.9%, 18/90) than in omnivores (2.2%, 2/90). Analysis of sequences showed that eight of the isolates - two from spotted hyaena (Crocuta crocuta) (2/8), three from lion (Panthera leo) (3/13), one from leopard (Panthera pardus) (1/6), one from small spotted genet (Genetta genetta) (1/2) and one Nile monitor lizard (Varanus niloticus) (1/2) - conformed to Trichinella zimbabwensis. One isolate from a hyaena was grouped under the encapsulated species clade comprising T. nelsoni and genotype Trichinella T8 reported to be present in South Africa. This is the first report confirming natural infection by T. zimbabwensis in hyaena, leopard, genet and Nile monitor lizard, adding to the body of knowledge on the epidemiology of Trichinella infections in the Greater Kruger National Park of South Africa. Ten Trichinella-like larval isolates recovered after digestion from four wildlife species in this study (2012-2016) revealed inconclusive results due to DNA degradation resulting from poor storage or too few larvae for analysis, in comparison to 20 unidentified isolates from five wildlife species during the 1964-2011 period.

The occurrence of Trichinella zimbabwensis in naturally infected wild crocodiles (Crocodylus niloticus) from the Kruger National Park, South Africa.

Trichinella zimbabwensis has been found naturally infecting crocodiles (Crocodylus niloticus) in Zimbabwe, Mozambique, Ethiopia and South Africa, as well as monitor lizards (Varanus niloticus) in Zimbabwe. The reports on natural infections were mostly accidental rather than structured surveys and involved very few animals. Previous surveillance studies in South Africa reported a 38.5% prevalence of T. zimbabwensis among wild crocodiles tested from the Mpumalanga province and Kruger National Park (KNP). No studies have been conducted to date on the geographical distribution and occurrence of T. zimbabwensis in wild crocodiles and varans in countries in southern Africa. Recent outbreaks of pansteatitis in crocodile populations of the KNP, South Africa, provided an opportunity to conduct a more structured survey aimed at elucidating the occurrence and distribution of T. zimbabwensis in culled wild crocodile populations within the KNP. Results from this study showed that T. zimbabwensis occurred in 10 out of 12 culled crocodiles form the KNP. The results also showed that the natural distribution of T. zimbabwensis in crocodiles includes all the major river systems in the KNP. The predilection sites of larvae in muscles followed a different pattern in naturally infected crocodiles compared to observations in experimentally infected mammalian hosts.

Trichinella zimbabwensis in a naturally infected mammal.

Trichinella zimbabwensis has been detected in wild and farmed Nile crocodiles (Crocodylus niloticus) and in wild monitor lizards (Varanus niloticus) of several African countries, but it has never been detected in mammals in nature, in spite of its infectivity to rodents, pigs, foxes and monkeys under laboratory conditions. The aim of this work was to describe the first detection of T. zimbabwensis in a naturally infected lion (Panthera leo) of the Kruger National Park (KNP) of South Africa. The sequence of the expansion segment V, a highly variable non-coding sequence of the large subunit ribosomal RNA of the genus Trichinella, of larvae from the lion was identical to that of larvae of T. zimbabwensis collected from a Nile crocodile originating from the same locality as the lion, suggesting a possible transmission of this parasite between mammals and reptiles. The KNP proves to be a very interesting area for parasites of the genus Trichinella since three taxa (Trichinella nelsoni, Trichinella T8 and T. zimbabwensis) circulate among the wildlife of this protected area.

Prevention by a silymarin/phospholipid compound of ethanol-induced social learning deficits in rats.

We explored the possibility that silymarin (SY), a fraction from Silybum marianum, might protect against the effects of in utero exposure to ethanol upon subsequent social memory function. Three groups of 8 pregnant female Sprague-Dawley rats each were provided with a liquid diet containing 35% ethanol derived calories (EDC). One experimental group received a daily subcutaneous injection of 400 mg/kg SY, the second, a 400 mg/kg oral dose of SY, a third group was maintained on the 35% EDC diet. A fourth group served as the pair-fed control group. The liquid diet regimen was maintained throughout pregnancy. Rats pups were fostered by dams in a fifth group that had been maintained on rat chow. At 90 days the pups were tested for social memory. Social recognition scores recorded for the ethanol pups were significantly poorer than those observed in both SY/ethanol groups and the chow group.

Protective effects of the flavonoid mixture, silymarin, on fetal rat brain and liver.

We investigated the possibility that the flavonoid mixture, silymarin (SY), administered as the compound Silymarin Phytosome (PHYTO), could protect the fetus from maternally ingested EtOH. Seventy-six female rats were randomly assigned to one of seven groups: pair-fed control; chow fed control; EtOH; and four groups receiving EtOH and PHYTO in varying dosages. All groups except the chow-fed control were maintained on a liquid diet. On day 1 of pregnancy the dams began the treatment protocol. On day 21 of pregnancy the rats were sacrificed and the fetuses removed. Gamma glutamyl transpeptidase (GGTP) activity was determined for liver and brain tissue from both the fetuses and the dams. GGTP activity in the EtOH/silymarin treatment groups did not differ significantly from that observed for the pair-fed control group. The observed GGTP activity levels for the EtOH-only group were significantly higher than those attained by the pair-fed control group. Although GGTP activity did not vary significantly with the quantity of PHYTO administered, as PHYTO dose was increased, GGTP activity decreased.

Female alcoholic outpatients and female college students: a correlational study of self-reported alcohol consumption and carbohydrate-deficient transferrin levels.

OBJECTIVE: We sought to test the efficacy of two biochemical markers of alcohol consumption, gamma glutamyl transpeptidase and carbohydrate deficient transferrin, as indicators of heavy alcohol consumption in a female population. METHOD: Using a sample of female outpatient alcoholics (n = 36) and a comparison group of female college students (n = 50), alcohol intake was monitored by self-report at approximately 90-day intervals over a period of 12 months. Gamma glutamyl transpeptidase (GGTP) and carbohydrate-deficient transferrin (CDT) values were determined for each sampling period. The criterion for heavy alcohol consumption was set at 140 g/alcohol/week for a 90-day period. Receiver operating characteristic (ROC) curves were plotted, area under the curve (Az) computed, and sensitivity and specificity calculated for both CDT and GGTP. ROC curves provide a graphical illustration of the association between the specificity and sensitivity of any diagnostic test over all possible cutpoint values. RESULTS: A significant, positive correlation between the amount of alcohol consumed and GGTP activity was recorded for the outpatient alcoholic group. Among the college students, there was a significant, positive correlation between CDT levels and alcohol consumed for the second reporting period. The sensitivity of the individual biochemical markers was low but, when used in combination, sensitivity was 66% and specificity, 80%. When a more stringent criterion for heavy alcohol consumption was applied (420 g/alcohol/week), the sensitivity of CDT actually decreased. CONCLUSIONS: CDT, used alone, is a poor indicator of heavy alcohol consumption in female subjects. When GGTP measures are used in conjunction with CDT, detection of heavy alcohol consumption may be enhanced to useful levels.

Carbohydrate-deficient transferrin levels in a female population.

Female college students (n = 49) from a small southwestern United States university participated in the 9-month study. Data collected included the assessment of drinking habits and other related substance use habits and serum levels of carbohydrate-deficient transferrin (CDT). Each subject provided an interview and blood sample on three occasions at 90-day intervals. Appended to the interview were a series of questions regarding stage of menstrual cycle and the diagnoses of certain diseases. The CDT values obtained were consistent with those obtained in other studies. Moderate-drinking subjects had significantly higher CDT values than did the abstainers and light drinkers. Females using oral contraceptives had significantly higher CDT values than those who were not taking oral contraceptives. Finally, although CDT values varied over time, they did not appear to vary as a function of menstrual cycle stage.

Alcohol consumption: biochemical and personality correlates in a college student population.

The frequency of alcohol use among a subject population of 28 male and 60 female college students was assessed using the Student Alcohol and Drug Use Survey (STADUS). Data were also collected on personality traits as measured by the Sensation Seeking Scale V (SSSV) and the Eysenck Personality Questionnaire (EPQ). Finally, three biochemical variables were assessed: monoamine oxidase (MAO) activity, dopamine beta hydroxylase (DBH) activity, and testosterone levels. Among males, high SSSV scores, high testosterone levels, and low MAO activity contributed to the variance in alcohol use, whereas among females, a significant proportion of the variability in alcohol use was accounted for by high SSSV scores, high DBH activity, and younger age.

A correlational study of carbohydrate-deficient transferrin values and alcohol consumption among Hispanic college students.

One hundred and forty-eight U.S. college students from a small southwestern university were asked to provide or complete the following: the Michigan Alcoholism Screening Test (MAST), a structured alcohol-use interview, and a 10-ml blood sample to be assayed for carbohydrate-deficient transferrin (CDT) levels and gamma-glutamyl-transpeptidase (GGTP) activity. Using the data obtained in the interview, only 2 females and 5 males were identified as heavy drinkers. Conclusions regarding the efficacy of the biochemical markers are therefore limited. It was found that CDT levels were significantly and positively correlated with various measures of alcohol consumption among males. There was no similar association between CDT, or GGTP, and alcohol consumption within the female sample.