Apoptosis is induced by sub-acute exposure to 3-MCPD and glycidol on Wistar Albino rat brain cells.

3-chloropropane-1,2-diol (3-MCPD) and its toxic metabolite glycidol were classified by the International Agency for Research on Cancer (IARC) as belonging to group 2B and 2A for humans. This study aimed to determine the sub-acute toxicity of these agents. Rats were exposed to 3-MCPD at 0.87 and 10 mg/kg/bw and glycidol (2,4 and 37,5 mg/kg/bw) for 90 days. miR-21 gene expression levels significantly decreased in all group's cerebellar tissues compared with control. Exposure to 10 mg/kg/bw 3-MCPD showed significant increases in PTEN in brain as compared to control group. The Akt gen expressions were significantly decreased in 3-MCPD and glycidol groups when compared to control group brains. Additionally, Caspase 3 and AIF immunopositivity significantly increased in 3-MCPD high dose and glycidol high dose groups in cerebellum granular layers compared to control. The results of the present study conclude that 3-MCPD and glycidol can induce apoptosis in rat brain tissue.

Efficacy of Azadirachta indica (Neem) leaf extract and hypertonic saline solution as intratesticular chemical sterilizing agents in dogs / Eficácia do extrato de folha de Azadirachta indica (Neem) e solução salina hipertônica como agente químico esterilizante intratesticular em cães

Castration refers to induced sterility via physical, chemical, or hormonal methods. Chemical castration is an efficient and reliable technique in contrast to other sterilization procedures as it is less painful to physical methods and cost­effective to hormonal methods. Azadirachta indica (neem), is a charismatic plant as its leaves possess anti­inflammatory, anti­microbial, and anti­androgenic chattels. To abate the escalating human population in South Asia, neem oil and neem leaf extract have been effectively used as a contraceptive agent. The key determinant of the current study was to evaluate Neem as a chemical sterilizing agent, (either necrotic or apoptotic), in dogs injected intratesticular in comparison to a hypertonic saline solution. Pre­ and post­injection testicular width size and blood samples for serum testosterone levels were collected on alternative days. Results disclosed substantial changes in testicular width size, histopathological profile, and serum testosterone level. A non­significant (P > 0.05) pre­injection testicular width readings in contrast to a significant increase (P < 0.05) three days post­injection was noted in all the competitive groups. The mean values recorded for testicular width size at the end of the trial study via neem leaf extract, 30% HSS and, control groups were 27.7362 ± 2.3315mm, 30.9594 ± 4.6861mm, and 24.5023 ± 2.5387mm, respectively. A declining trend, regarding serum testosterone level being statistically significant (P < 0.05) was recorded in treated groups (A, B) in contrast to the control group (C) as the values were 1.5357 ± 0.7819ng, 1.2669 ± 0.9095ng, and 2.4517 ± 0.1827ng in groups A, B, and C, respectively. Histopathological findings advocated the presence of apoptotic bodies in the neem treated group whereas the presence of degenerated interstitial cells, necrosed seminiferous tubules, damaged germinal epithelium, and ceased spermatogenesis was also studied in both competitive groups. Thus, the apoptotic effect and anti­inflammatory property of neem leaf extract resulted in less painful castration and verified Azadirachta indica as a better substitute for chemical castration in contrast to hypertonic saline solution.(AU)

A castração consiste na indução da esterilidade por meio físico, químico ou hormonal. A castração química é uma técnica eficiente e confiável, em contraste com outros procedimentos de esterilização, pois é menos dolorosa para os métodos físicos e econômicos para os métodos hormonais. Azadirachta indica (neem), é uma planta carismática, pois possui folhas anti­inflamatórias, antimicrobianas e antiandrogênicas. Para diminuir a crescente população humana no sul da Ásia, o óleo de nim e o extrato de folhas de nim têm sido efetivamente usados como agente contraceptivo. O principal determinante deste estudo atual foi avaliar o Neem como um agente esterilizante químico (necrótico ou apoptótico) em cães injetados intratesticularmente em comparação com uma solução salina hipertônica. O tamanho da largura testicular pré e pós­injeção e as amostras de sangue para os níveis séricos de testosterona foram colhidas em dias alternados. Os resultados obtidos revelaram alterações substanciais no tamanho da largura testicular, perfil histopatológico e nível sérico de testosterona. Observou­se uma leitura não significativa (P> 0,05) da largura testicular da pré­injeção, em contraste com um aumento significativo (P <0,05) três dias após a injeção em todos os grupos competitivos.Os valores médios registrados para o tamanho da largura testicular no final do estudo via extrato de folhas de nim,HSS a 30% e grupos controle foram 27,7362 ± 2,3315 mm, 30,9594 ± 4,6861 mm e 24,5023 ± 2,5387 mm, respectivamente.Uma tendência decrescente, com relação ao nível sérico de testosterona sendo estatisticamente significante (P <0,05), foi registrada nos grupos tratados (A, B), em contraste com o grupo controle (C), pois os valores eram 1,5357 ± 0,7819ng, 1,2669 ± 0,9095ng e 2,4517 ± 0,1827ng nos grupos A, B e C, respectivamente. Os achados histopatológicos advogaram a presença de corpos apoptóticos no grupo tratado com nim, enquanto a presença de células intersticiais degeneradas, túbulos seminíferos necrosados, epitélio germinativo danificado e espermatogênese interrompida também foi estudada nos dois grupos competitivos. Assim, o efeito apoptótico e a propriedade anti­inflamatória do extrato de folhas de nim resultaram em uma castração menos dolorosa e confirmaram que a Azadirachta indica foi um melhor substituto para a castração química do que a solução salina hipertônica.(AU)

DNA methylation analysis in rat kidney epithelial cells exposed to 3-MCPD and glycidol.

3-Monochloropropane-1,2-diol (3-MCPD) is a well-known food processing contaminant that has been regarded as a rat carcinogen, which is known to induce Leydig-cell and mammary gland tumors in males, as well as kidney tumors in both genders. 3-MCPD is highly suspected to be a non-genotoxic carcinogen. 2,3-Epoxy-1-propanol (glycidol) can be formed via dehalogenation from 3-MCPD. We aimed to investigate the cytotoxic effects of 3-MCPD and glycidol, then to demonstrate the possible epigenetic mechanisms with global and gene-specific DNA methylation in rat kidney epithelial cells (NRK-52E). IC50 value of 3-MCPD was determined as 48 mM and 41.39 mM, whereas IC50 value of glycidol was 1.67 mM and 1.13 mM by MTT and NRU test, respectively. Decreased global DNA methylation at the concentrations of 100 µM and 1000 µM for 3-MCPD and 100 µM and 500 µM for glycidol were observed after 48 h exposure by using 5-methylcytosine (5-mC) ELISA kit. Methylation changes were detected in promoter regions of c-myc and Rassf1a in 3-MCPD and glycidol treated NRK-52E cells by using methylation-specific PCR (MSP), whereas changes on gene expression of c-myc and Rassf1a were observed by using real-time PCR. However, e-cadherin, p16, VHL and p15 genes were unmethylated in their CpG promoter regions in response to treatment with 3-MCPD and glycidol. Alterations in DNA methylation might be key events in the toxicity of 3-MCPD and glycidol.

Effect of piglet castration with nonsurgical sterilant on the zootechnical performance and pork carcass quality / Efeito da castração de leitões com esterilizante não cirúrgico no desempenho zootécnico e na qualidade da carcaça suína

A non-surgical sterilant (NSS) was used with the objective of assessing the zootechnical performance, carcass and meat quality, and hormone levels. 90 male piglets were selected with birthweights varying between 1.5kg to 2.0kg. The control group was constituted of 45 males castrated by the conventional surgical method, at the seventh day of age, and the treated group was constituted of 45 males castrated with NSS (active principle of zinc gluconate in the concentration of 26.2mg/mL associated to the dimethyl sulfoxide at 0.5%), with the application of the first dose on the seventh day of age and the second dose on the fourteenth day of age. The zootechnical assessments were carried out on the farm and consisted: weight gain in the periods per animal and feed conversion. The slaughter of the animals and the assessment of the carcass and meat quality and hormone levels were carried out in a slaughterhouse and the municipality of Campinas (SP). The study demonstrated that the use of NSS was a viable alternative in relation to the zootechnical performance and the carcass quality. In relation to the meat quality there was no significant difference in the majority of the assessed parameters.(AU)

Foi utilizado esterilizante não cirúrgico (ENC) com o objetivo de se avaliar o desempenho zootécnico e a qualidade de carcaça, de carne e de níveis hormonais. Foram selecionados 90 leitões machos de peso entre 1,5kg e 2,0kg ao nascimento. O grupo controle foi constituído por 45 machos castrados pelo método cirúrgico convencional, ao sétimo dia de idade, e o grupo tratado foi constituído por 45 machos castrados com ENC (princípio ativo de gluconato de zinco na concentração de 26,2mg/mL associado ao dimetil sulfóxido a 0,5%), com primeira dose de aplicação ao sétimo dia de idade e a segunda dose ao 14° dia de idade. As avaliações consistiram de: ganho de peso nos períodos por animal e conversão alimentar. O abate dos animais e a avaliação da carne e de níveis hormonais foram realizados em um frigorífico, na cidade de Campinas (SP). O uso de ENC foi uma alternativa viável em relação ao desempenho zootécnico e à qualidade da carcaça. Em relação à qualidade da carne, não houve diferença significativa na maioria dos parâmetros avaliados.(AU)

Chemosterilants for Control of Insects and Insect Vectors of Disease.

Both historically and at present, vector control is the most generally effective means of controlling malaria transmission. Insecticides are the predominant method of vector control, but the sterile insect technique (SIT) is a complementary strategy with a successful track record in both agricultural and public health sectors. Strategies of genetic and radiation-induced sterilization of Anopheles have to date been limited by logistical and/or regulatory hurdles. A safe and effective mosquito chemosterilant would therefore be of major utility to future deployment of SIT for malaria control. Here we review the prior and current use of chemosterilants in SIT, and assess the potential for future research. Recent genomic and proteomic studies reveal opportunities for specific targeting of seminal fluid proteins, and the capacity to interfere with sperm motility and storage in the female.

3-Monochloro-1,2-propanediol (3-MCPD) induces apoptosis via mitochondrial oxidative phosphorylation system impairment and the caspase cascade pathway.

3-Monochloro-1,2-propanediol (3-MCPD) is the most toxic chloropropanols compounds in foodstuff which mainly generated during thermal processing. Kidney is one of the primary target organs for 3-MCPD. Using human embryonic kidney cell (HEK293FT) as an in vitro model, we found that 3-MCPD caused concentration-dependent increase in cytoxicity as assessed by dye uptake, lactatedehydrogenase (LDH) leakage and MTT assays. HEK293FT cell treated with 3-MCPD suffered the decrease of mitochondrial membrane potential and the impairment of mitochondrial oxidative phosphorylation system, especially the reduced amount of mRNA expression and protein synthesis of electron transport chain complex II, complex IV, and complex III. More importantly, energy release (ATP synthesis) was significantly inhibited by 3-MCPD resulting from the down regulation expressions of ATP synthase (ATP6 and ATP8), as well as the loss of transmembrane potential required for synthesis of ATP. The decreased ratio of mitochondrial apoptogenic factors Bax/Bcl-2 and the cytochrome-c release from mitochondria to cytosol followed by the activation of apoptotic initiators caspase 9 and apoptotic executioners (caspase 3, caspase 6 and caspase 7) leading to apoptosis. The activation of caspase 8 and caspase 2 implied that there were probably other factors to induce the caspase-dependent apoptosis.

Dihydroisoxazole inhibitors of Anopheles gambiae seminal transglutaminase AgTG3.

BACKGROUND: Current vector-based malaria control strategies are threatened by the rise of biochemical and behavioural resistance in mosquitoes. Researching mosquito traits of immunity and fertility is required to find potential targets for new vector control strategies. The seminal transglutaminase AgTG3 coagulates male Anopheles gambiae seminal fluids, forming a 'mating plug' that is required for male reproductive success. Inhibitors of AgTG3 can be useful both as chemical probes of A. gambiae reproductive biology and may further the development of new chemosterilants for mosquito population control. METHODS: A targeted library of 3-bromo-4,5-dihydroxoisoxazole inhibitors were synthesized and screened for inhibition of AgTG3 in a fluorescent, plate-based assay. Positive hits were tested for in vitro activity using cross-linking and mass spectrometry, and in vivo efficacy in laboratory mating assays. RESULTS: A targeted chemical library was screened for inhibition of AgTG3 in a fluorescent plate-based assay using its native substrate, plugin. Several inhibitors were identified with IC50 < 10 µM. Preliminary structure-activity relationships within the library support the stereo-specificity and preference for aromatic substituents in the chemical scaffold. Both inhibition of plugin cross-linking and covalent modification of the active site cysteine of AgTG3 were verified. Administration of an AgTG3 inhibitor to A. gambiae males by intrathoracic injection led to a 15% reduction in mating plug transfer in laboratory mating assays. CONCLUSIONS: A targeted screen has identified chemical inhibitors of A. gambiae transglutaminase 3 (AgTG3). The most potent inhibitors are known inhibitors of human transglutaminase 2, suggesting a common binding pose may exist within the active site of both enzymes. Future efforts to develop additional inhibitors will provide chemical tools to address important biological questions regarding the role of the A. gambiae mating plug. A second use for transglutaminase inhibitors exists for the study of haemolymph coagulation and immune responses to wound healing in insects.

Effects of neem oil (Azadirachta indica A. Juss) on midgut cells of predatory larvae Ceraeochrysa claveri (Navás, 1911) (Neuroptera: Chrysopidae).

The effects of ingested neem oil, a botanical insecticide obtained from the seeds of the neem tree, Azadirachta indica, on the midgut cells of predatory larvae Ceraeochrysa claveri were analyzed. C. claveri were fed on eggs of Diatraea saccharalis treated with neem oil at a concentration of 0.5%, 1% and 2% during throughout the larval period. Light and electron microscopy showed severe damages in columnar cells, which had many cytoplasmic protrusions, clustering and ruptured of the microvilli, swollen cells, ruptured cells, dilatation and vesiculation of rough endoplasmic reticulum, development of smooth endoplasmic reticulum, enlargement of extracellular spaces of the basal labyrinth, intercellular spaces and necrosis. The indirect ingestion of neem oil with prey can result in severe alterations showing direct cytotoxic effects of neem oil on midgut cells of C. claveri larvae. Therefore, the safety of neem oil to non-target species as larvae of C. claveri was refuted, thus the notion that plants derived are safer to non-target species must be questioned in future ecotoxicological studies.

Effects of quinestrol on reproductive hormone expression, secretion, and receptor levels in female Mongolian gerbils (Meriones unguiculatus).

Quinestrol, a synthetic estrogen with marked estrogenic effects and prolonged activity, has potential as a contraceptive for Mongolian gerbils. The objective of this study was to describe the effects of quinestrol on reproductive hormone expression, secretion, and receptor levels in female Mongolian gerbils. Serum and pituitary concentrations of follicle stimulating hormone (FSH) and luteinizing hormone (LH) were decreased, whereas serum concentrations of estradiol (E2) and progesterone (P4) were increased after quinestrol treatment; the effects were both time- and dose-dependent. Furthermore, quinestrol downregulated expression of FSHß and LHß mRNA in the pituitary gland, as well as FSH receptor (FSHR) and estrogen receptor (ER) ß in the ovary. However, it up-regulated mRNA expression levels of ERα and progesterone receptor (PR) in the pituitary gland and uterus, as well as mRNA for LH receptor (LHR) and PR in the ovary (these effects were time- and dose-dependent). In contrast, quinestrol had no significant effects on the mRNA expression levels of ERα in the ovary, or the gonadotropin α (GtHα) subunit in the pituitary gland. We inferred that quinestrol impaired synthesis and secretion of FSH and LH and that the predominant ER subtype in the pituitary gland of Mongolian gerbils may be ERα. Overall, quinestrol disrupted reproductive hormone receptor expression at the mRNA level in the pituitary-gonadal axis of the Mongolian gerbil.

Evaluating the anti-fertility potential of α-chlorohydrin on testis and spermatozoa in the adult male wild Indian house rat (Rattus rattus).

To examine the effects of α-chlorohydrin on testis and cauda epididymis in the male house rat (Rattus rattus), 24 adult male rats were segregated into two groups. Group I rats were force-fed daily by intragastric intubation with α-chlorohydrin at a single dose of 1.0 mg/100 g body weight/d for 5, 15, and 45 days. Another group was fed with distilled water, which served as the control. The treated male rats were paired with 24 adult proestrus female rats for 5 days after the last oral treatment and fertility was tested. At the end of the experiments, all of the male rats were weighed and killed by cervical dislocation. The right testes were removed, weighed, and processed for ultrastructural changes of spermatozoa from the cauda epididymis and testis under scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The seminiferous tubular area, nuclear diameter of the Sertoli and Leydig cells, percentage of spermatogonia, primary spermatocytes, secondary spermatocytes, spermatids, spermatozoa, and Sertoli cells in each group were compared morphometrically. Our results showed that the percentages of primary spermatocytes steadily increased from 5 to 15 days, but primary and secondary spermatocytes decreased significantly at 45 days. There was a steady decline in the percentages of spermatozoa and spermatids at all fixation intervals in the treated animals, but the percentages of spermatogonia and Sertoli cells increased significantly at 15 and 45 days. Seminiferous tubular areas, nuclear diameter of Leydig and Sertoli cells, and fertility rates were reduced after 45 days of treatment. SEM and TEM studies revealed severe morphological abnormalities in the spermatozoa, including deglutination of the acrosomal part, loss of head capsules, and fragmentation of tail fibrils. There was an enhanced anti-fertility effect and a lower number of implantation sites in the rats treated for 5 days. Our results validate α-chlorohydrin as a successful anti-fertility agent that prevents spermatogenesis.

Proposta de diretrizes sanitárias para avaliação do prazo de validade de saneantes com ação antimicrobiana de uso na assistência à saúde / Proposal of sanitary lines of direction for evaluation of the shelf life of health assistance biocides

Até a finalização deste estudo, não existia no Brasil uma legislação no seguimento de produtos saneantes que disponibilizasse um guia cientificamente embasado para realização de estudos de estabilidade. O objetivo deste trabalho consiste na análise das legislações para registro de produtos saneantes de uso em assistência à saúde no Brasil, comparando seus diferentes critérios de estudo com legislações internacionais. Como resultado desta análise, propõem-se um guia para garantir a eficácia e a segurança ao longo do prazo de validade definido para estes produtos. Verifica-se que as legislações internacionais estudadas apresentam critérios mais rígidos, tais como (i) a avaliação das especificações de qualidade e segurança ao longo de todo o prazo de validade proposto; (ii) a avaliação do comportamento da embalagem e informações da rotulagem; (iii) o uso de uma metodologia de análise validada; (iv) e quando aplicável um estudo que simule as variações do produto nas condições de uso (ex.: diluições ou ativação para uso). Após análise crítica destas legislações, chega-se à proposta de um guia adequado às realidades do setor produtivo brasileiro, de forma a se regulamentar este procedimento como parte integrante do processo administrativo de registro sanitário nas rotinas da Agencia Nacional de Vigilância Sanitária - Brasil (ANVISA).

Estabelecimento de protocolo para micropropagação de Lippia gracilis Schauer / Establishment of protocol for Lippia gracilis Schauer micropropagation

A espécie Lippia gracilis SCHAUER (Verbenaceae) é nativa do Nordeste brasileiro e se destaca pela capacidade de acumular nos tricomas glandulares óleos essenciais com atividade antimicrobiana. Tendo em vista que não constam trabalhos na literatura sobre o estabelecimento in vitro dessa espécie, este trabalho teve como objetivo estabelecer protocolo para micropropagação de L. gracilis. Para tanto, ramos contendo folhas foram coletados de plantas matrizes no habitat natural para a confecção de estacas. Em laboratório, os explantes provenientes do processo de estaquia foram assepticamente tratados e inoculados em meio MS, acrescido de fitorreguladores, com o intuito de se estabelecer a melhor dose para o desenvolvimento dos explantes. Em decorrência de altos níveis de contaminação, avaliou-se o efeito da cefalexina. No entanto, apesar do antibiótico ter apresentado diminuição na contaminação bacteriana, a porcentagem de oxidação foi elevada. Portanto, testou-se o carvão ativado, ácido ascórbico, ácido cítrico e metade dos sais de MS quanto a eficiência no controle da oxidação. Concluiu-se que, o antibiótico na concentração utilizada provocou a oxidação dos explantes e os fitorreguladores, bem como os métodos antioxidantes testados, não apresentaram resultados consistentes para o melhor desenvolvimento dos explantes e controle da oxidação, respectivamente.

The species Lippia gracilis Schauer (Verbenaceae) is native to Northeastern Brazil and has been important for its ability to accumulate essential oils with antimicrobial activity in the glandular trichomes. Since there are no reports in the literature on the micropropagation of this species, the present work aimed to establish a protocol for L. gracilis micropropagation. Thus, branches containing leaves were collected from plant matrices in their natural habitat to prepare cuttings. In the laboratory, explants from cutting were aseptically treated and inoculated onto MS medium plus plant growth regulators in order to establish the best dose for the development of explants. Due to high levels of contamination, the effect of cephalexin was evaluated. Although the antibiotic decreased the bacterial contamination, the percentage of oxidation was high. Then, activated charcoal, ascorbic acid, citric acid and half the salts of MS were tested for their effectiveness to control oxidation. In conclusion, the used antibiotic concentration resulted in oxidation of explants. Furthermore, plant growth regulators and antioxidant methods did not show consistent results for a better development of explants and control of oxidation, respectively.

[Potential of Metarhizium anisopliae and Beauveria bassiana isolates and neem oil to control the Aphid Lipaphis erysimi (Kalt.) (Hemiptera: Aphididae)]. / Potencial de isolados de Metarhizium anisopliae e Beauveria bassiana e do óleo de Nim no controle do pulgão Lipaphis erysimi (Kalt.) (Hemiptera: Aphididae).

This work aimed to determine the efficiency of the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana to control the aphid Lipaphis erysimi (Kalt.) (Hemiptera: Aphididae) in kale Brassica oleracea var acephala D.C., as well as their compatibility with a neem oil formulation (Neemseto). Ten isolates of both fungi were tested and the most pathogenic ones were B. bassiana CG001 and M. anisopliae CG30 with 90% and 4.4 days, and 64% and 3.8 days of mortality and median lethal time, respectively. Bioassays with neem at concentrations of 0.5, 1.0 and 2.0% were done either by leaf discs dipping or spraying the aphids on the leaf discs. The neem spraying treatment at 2.0% provided 90% mortality. The use of B. bassiana isolate CG001 or M. anisopliae isolate CG30 with neem at 0.125, 0.25, and 0.5%, demonstrated that these isolates could have their spore viability or colony growth affected when exposed to neem concentrations higher than 0.25%. In absolute values, the isolates B. bassiana CG001 and M. anisopliae CG30 are the most virulent to L. erysimi, and could be utilized in the management of this pest.

Potencial de isolados de Metarhizium anisopliae e Beauveria bassiana e do óleo de Nim no controle do pulgão Lipaphis erysimi (Kalt. ) (Hemiptera: Aphididae) / Potential of Metarhizium anisopliae and Beauveria bassiana isolates and Neem oil to control the Aphid Lipaphis erysimi (Kalt. ) (Hemiptera: Aphididae)

Avaliou-se a eficiência de Metarhizium anisopliae e Beauveria bassiana para o controle do pulgão Lipaphis erysimi (Kalt.) em couve Brassica oleracea L. var. acephala D.C., bem como sua compatibilidade com óleo de nim (Neemseto®). Dez isolados desses fungos foram utilizados, onde o isolado CG 001 de B. bassiana e o isolado CG 30 de M. anisopliae apresentaram-se como os mais virulentos com 90 por cento e 4,4 dias, e 64 por cento e 3,8 dias, de mortalidade e tempo letal médio, respectivamente. Bioensaios com o produto à base de nim nas concentrações 0,5; 1,0 e 2,0 por cento foram realizados por imersão foliar e pulverização sobre os pulgões. O tratamento com pulverização de 2,0 por cento de Neemseto® proporcionou mortalidade de 90 por cento. O teste in vitro de Neemseto® a 0,125; 0,25 e 0,5 por cento, sobre os isolados CG 001 de B. bassiana e CG 30 de M. anisopliae mostrou que esses isolados podem ter seu crescimento colonial e viabilidade alterados quando expostos a concentrações de nim maiores que 0,25 por cento. Em valores absolutos, os isolados CG 001 de B. bassiana e CG 30 de M. anisopliae foram os mais virulentos para L. erysimi, podendo ser utilizados no manejo dessa praga.

This work aimed to determine the efficiency of the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana to control the aphid Lipaphis erysimi (Kalt.) (Hemiptera: Aphididae) in kale Brassica oleracea var acephala D.C., as well as their compatibility with a neem oil formulation (Neemseto®). Ten isolates of both fungi were tested and the most pathogenic ones were B. bassiana CG001 and M. anisopliae CG30 with 90 percent and 4.4 days, and 64 percent and 3.8 days of mortality and median lethal time, respectively. Bioassays with neem at concentrations of 0.5, 1.0 and 2.0 percent were done either by leaf discs dipping or spraying the aphids on the leaf discs. The neem spraying treatment at 2.0 percent provided 90 percent mortality. The use of B. bassiana isolate CG001 or M. anisopliae isolate CG30 with neem at 0.125, 0.25, and 0.5 percent, demonstrated that these isolates could have their spore viability or colony growth affected when exposed to neem concentrations higher than 0.25 percent. In absolute values, the isolates B. bassiana CG001 and M. anisopliae CG30 are the most virulent to L. erysimi, and could be utilized in the management of this pest.

Sperm mitochondrial integrity is not required for hyperactivated motility, zona binding, or acrosome reaction in the rhesus macaque.

Whether the main energy source for sperm motility is from oxidative phosphorylation or glycolysis has been long-debated in the field of reproductive biology. Using the rhesus monkey as a model, we examined the role of glycolysis and oxidative phosphorylation in sperm function by using alpha-chlorohydrin (ACH), a glycolysis inhibitor, and pentachlorophenol (PCP), an oxidative phosphorylation uncoupler. Sperm treated with ACH showed no change in percentage of motile sperm, although sperm motion was impaired. The ACH-treated sperm did not display either hyperactivity- or hyperactivation-associated changes in protein tyrosine phosphorylation. When treated with PCP, sperm motion parameters were affected by the highest level of PCP (200 microM); however, PCP did not cause motility impairments even after chemical activation. Sperm treated with PCP were able to display hyperactivity and tyrosine phosphorylation after chemical activation. In contrast with motility measurements, treatment with either the glycolytic inhibitor or the oxidative phosphorylation inhibitor did not affect sperm-zona binding and zona-induced acrosome reaction. The results suggest glycolysis is essential to support sperm motility, hyperactivity, and protein tyrosine phosphorylation, while energy from oxidative phosphorylation is not necessary for hyperactivated sperm motility, tyrosine phosphorylation, sperm-zona binding, and acrosome reaction in the rhesus macaque.

Occurrence of chloropropanols in soy sauce and other foods in China between 2002 and 2004.

A survey of chloropropanols in soy sauce and some selected foods in China is reported. Thirty-seven traditionally brewed soy sauce samples contained 3-MCPD below the EC maximum limit (ML) of 0.02 mg kg(-1). All soy sauce samples (629) from retailers contained levels of 3-MCPD ranging between <0.005 (LOQ) and 189 mg kg(-1), and only 12.2% had levels in excess of the Chinese ML of 1.0 mg kg(-1) for acid hydrolyzed vegetable protein (acid HVP). This indicates that the necessary processing changes have been made to decrease levels of chloropropanols in soy sauce. 2-Monochloropropane-1,3-diol (2-MCPD), 1,3-dichloro-2-propanol (1,3-DCP) and 2,3-dichloro-1-propanol (2,3-DCP) were detected in 48.1 19.1 and 3.78% of the soy sauce samples, respectively; the highest levels being 20.3, 8.26 and 0.50 mg kg(-1), respectively. A good linear correlation was found between the amount of 3-MCPD and 2-MCPD, with the level of 3-MCPD being generally higher than that of other chloropropanols for the same soy sauce. Acid HVP contained 3-MCPD at a level of 0.010-117.7 mg kg(-1) (on a liquid basis) and 80% of samples contained levels exceeding 1.0 mg kg(-1). In some other foods investigated, relatively high levels of 3-MCPD were found in soy sauce powder, oyster sauce, beef products, instant noodle spices and health foods, ranging from 0.029 to 13.64 mg kg(-1). It is concluded that abnormal levels of 3-MCPD in soy sauce or other foods produced in China may result from acid hydrolysis or the addition of the contaminated acid HVP.