The entomopathogenic Autographa californica (Speyer) nucleopolyhedrovirus (AcMNPV) has been genetically modified to increase its speed of kill. The potential adverse effects of a recombinant AcMNPV (AcAaIT) as well as wild type AcMNPV and wild type Spodoptera littoralis NPV (SlNPV) were studied. Cotton plants were treated with these viruses at concentrations that were adjusted to resemble the recommended field application rate (4 x 10¹² PIBs/feddan, feddan = 4,200 m²) and 3^rd instar larvae of S. littoralis were allowed to feed on the contaminated plants. SDS-PAGE, ELISA, and DNA analyses were used to confirm that larvae that fed on these plants were virus-infected. Polyhedra that were purified from the infected larvae were subjected to structural protein analysis. A 32 KDa protein was found in polyhedra that were isolated from all of the viruses. Subtle differences were found in the size and abundance of ODV proteins. Antisera against polyhedral proteins isolated from AcAaIT polyhedra were raised in rabbits. The terminal bleeds from rabbits were screened against four coating antigens (i.e., polyhedral proteins from AcAaIT, AcAaIT from field-infected larvae (AcAaIT-field), AcMNPV, and SlNPV) using a two-dimensional titration method with the coated antigen format. Competitive inhibition experiments were conducted in parallel to optimize antibody and coating antigen concentrations for ELISA. The IC50 values for each combination ranged from 1.42 to 163 μg/ml. AcAaIT-derived polyhedrin gave the lowest IC50 value, followed by those of SlNPV, AcAaIT-field, and AcMNPV. The optimized ELISA system showed low cross reactivity for AcMNPV (0.87%), AcAaIT-field (1.2%), and SlNPV (4.0%). Genomic DNAs isolated from AcAaIT that were passaged in larvae of S. littoralis that were reared in the laboratory or field did not show any detectable differences. Albino rats (male and female) that were treated with AcAaIT, AcMNPV or SlNPV (either orally or by intraperitoneal injection at doses of 1 x 10⁸ or 1 x 10⁷ PIBs/rat, respectively) appeared to be healthy and showed increased body weight at 21 days posttreatment. The effect of virus administration on hematological, serum biochemical, and histopathological parameters were determined. Slight to moderate differences were observed in most of the hematological parameters. Specifically, serum proteins were decreased markedly in female rats treated orally with SlNPV, and in male rats injected with AcAaIT. SDS-PAGE analysis also showed some changes in serum protein profiles. No marked changes in acetylcholine esterase (AChE) activity were found. Changes in serum glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), bilirubin, creatinin, and urea were also observed. Immunohistochemical observation of tissues from stomach, intestine, liver, kidney, brain, spleen, and lung also showed slight changes. Fish (Tilapia nilotica) were also exposed to AcAaIT, AcMNPV or SlNPV by incorporating each of the viruses into diet (1 x 10⁹ PIBs/group). No mortality was found in treated or untreated fish during the experimental period (28 days). Macrophage phagocytic activity of fish head kidney cells increased with time, reaching maximum values at 180 min for both treated and control fish. Full article

Menia El-Kamh province of the Sharkia Governorate constitutes one of the largest agricultural areas in Egypt. About 88% of the nearly 472,000 people living in this province rely on agricultural activities for subsistence. Several pesticides including organochloride, organophosphorus, carbamate, and pyrethroid insecticides, fungicides, and herbicides are commonly used in citrus, vegetable and other crop-growing areas to increase agricultural productivity. However, their use has also been associated with several cases of pesticide poisoning. In this research, we conducted a field survey to assess the knowledge, attitudes, and practices of the farmer’s community regarding the safe use of pesticides. We also evaluated the residual concentrations of selected pesticides in water, soil, milk, fish, and orange samples, and estimated the potential health risks associated with the exposure to these pesticides. Data obtained from the field survey indicate that more than 95% of farm workers do not practice safety precautions during pesticide formulation and application; leading to a considerable prevalence of pesticide-related illnesses in this agricultural community. Pesticide residues in various environmental samples varied greatly; from below detection levels (3-5 ng) to as high as 325 ppb depending on the matrix of interest, and the specific pesticide of concern. The analysis of health risk estimates indicated that chlorpyrifos, DDT, dimethoate, methomyl, and larvin did not pose a direct hazard to human health, although present in water, milk, orange, and/or fish. However, aldicarb, and carbosulfan levels exceeded the reference doses, indicating a great potential for systemic toxicity, especially in children who are considered to be the most vulnerable population subgroup. The upper-bound values of cancer risk from DDT exposure were estimated to be about 8 (adults), and 55 (children) excess cancers in a population of one million. Full article