A sub-chronic feeding study of dual toxin insect-resistant transgenic maize (CEMB-413) on Wistar rats.

Genetically modified (GM) crops expressing insecticidal crystal proteins are widely accepted worldwide, but their commercial utilization demands comprehensive risk assessment studies. A 90-day risk assessment study was conducted on Wistar rats fed with GM maize (CEMB-413) expressing binary insect-resistant genes (cry1Ac and cry2Ab) at low (30%) and high (50%) dose along with a control diet group. The study used fifty Wistar rats randomly distributed in five treatment groups. Our study revealed that compared to controls, GM diet had no adverse effects on animal's health, including body weight, food consumption, clinical pathological parameters, serum hormone levels and histological parameters of testes and ovaries of rats. Differences were observed in transcripts levels of fertility related genes, but these were independent of treatment with GM diet.

Evaluation the effect of subchronic feeding of transgenic cotton line (CKC1) on the faecal microbiota of albino rabbits.

Recent studies have demonstrated a strong relationship between the intestinal microbiota and the host health. As such, consumers are increasingly becoming more concerned about the potential effect of certain foods/feeds, particularly of transgenic origin on the gut microbiota. Although the European Food Safety Authority has recommended in their guidelines, to study the effect of transgenic food/feed on host-microbiota, yet, few studies have focused on the evaluation of such effects mainly due to culturing difficulties. Therefore, this study was intended to evaluate the potential adverse effects of transgenic diet consumption on some specific gut microflora (Lactobacillus group, Bifidobacterium genus, Escherichia coli subgroup and Enterococcus genus) of rabbits. A total of forty-eight rabbits were randomly assigned into four groups and fed a diet containing a variable proportion of transgenic cottonseeds at 0, 20, 30 and 40% inclusion level, respectively. Changes in the specific or total faecal bacterial population were monitored at five different experimental stages (i.e. 0, 45, 90, 135 and 180 days) using both the traditional plate count method (TM) and quantitative real-time PCR (qPCR). No significant differences (p > .05) were observed concerning numbers of specific bacteria or total bacteria between the control and experimental groups, though qPCR showed numerically higher values in terms of 16S rRNA gene copies as compared to the values obtained from TM. However, such numerical differences were biologically insignificant (p > .05). Similarly, no significant variations were noticed in the calculated B/E (log10 copies of Bifidobacterium per g faces/log10 copies of E. coli genome per g faeces) ratios in all the groups. All the ratios were in the range of 1.24 to 1.30 throughout the experiment, indicating a good balance of intestinal microflora and greater resistance to intestinal disorders. It is therefore concluded that feeding transgenic cottonseeds could not adversely affect the gut microflora of rabbits during a long-term study.

A 90-day subchronic toxicity study of transgenic cotton expressing Cry1Ac, Cry2A and CP4-EPSPS proteins in Sprague-Dawley rats.

Genetically engineered crops expressing insecticidal and herbicide-tolerant traits offer a new strategy for crop protection and enhanced production; however, at the same time present a challenge in terms of toxicology and safety. The current experiment presents the findings of a 90-day feeding study in Sprague-Dawley rats with transgenic cottonseed which is expressing insecticidal Cry proteins (Cry1Ac and Cry2A), and tolerant to the herbicide glyphosate. There were 100 rats in this experiment divided into 5 groups of 10 rats/sex/group. Cottonseed from transgenic and control (near-isogenic) lines was formulated into standard diets at levels of 10% and 30% (w/w). All formulated diets were nutritionally balanced. Overall appearance, feed consumption, body weight, organ weight, haematology, serum chemistry and urinalysis were comparable between control and treatment groups. In addition, there was no treatment-related difference in findings of microscopic histopathology and gross appearance of tissues. In conclusion, following the 13-week of feeding transgenic cottonseed, no treatment-related adverse effects were observed in any of the parameters measured in this experiment. Thus, this study demonstrated that transgenic cottonseeds do not cause toxicity and are nutritionally equivalent to its conventional counterpart.

Comparative analysis of Constitutive and fiber-specific promoters under the expression pattern of Expansin gene in transgenic Cotton.

Promoters are specified segments of DNA that lead to the initiation of transcription of a specific gene. The designing of a gene cassette for plant transformation is significantly dependent upon the specificity of a promoter. Constitutive Cauliflower mosaic virus promoter, CaMV35S, due to its developmental role, is the most commonly used promoter in plant transformation. While Gossypium hirsutum (Gh) being fiber-specific promoter (GhSCFP) specifically activates transcription in seed coat and fiber associated genes. The Expansin genes are renowned for their versatile roles in plant growth. The overexpression of Expansin genes has been reported to enhance fiber length and fineness. Thus, in this study, a local Cotton variety was transformed with Expansin (CpEXPA1) gene, in the form of two separate cassettes, each with a different promoter, named as 35SEXPA1 and FSEXPA1 expressed under CaMV35S and GhSCFP promoters respectively. Integration and Spatiotemporal relative expression of the transgene were studied in an advanced generation. GhSCFP bearing transgene expression was significantly higher in Cotton fiber than other plant parts. While transgene with CaMV35S promoter was found to be continually expressing in all tissues but the expression was lower in fiber than that expressed under GhSCFP. The temporal expression profile was quite interesting with a gradual increasing pattern of both constructs from 1DPA (days post anthesis) to 18DPA and decreased expression from 24 to 30 DPA. Besides the relative expression of promoters, fiber cellulose quantification and fluorescence intensity were also observed. The study significantly compared the two most commonly used promoters and it is deduced from the results that the GhSCFP promoter could be used more efficiently in fiber when compared with CaMV35S which being constitutive in nature preferred for expression in all parts of the plant.

Assessing the fate of recombinant plant DNA in rabbit's tissues fed genetically modified cotton.

Various feeding studies have been conducted with the different species of animals to evaluate the possible transfer of transgenic DNA (tDNA) from genetically modified (GM) feed into the animal tissues. However, the conclusions drawn from most of such studies are sometimes controversial. Thus, in the present study, an attempt has been made to evaluate the fate of tDNA in rabbits raised on GM cotton-based diet through PCR analysis of the DNA extracted specifically from blood, liver, kidney, heart and intestine (jejunum). A total of 48 rabbits were fed a mixed diet consisting variable proportions of transgenic cottonseeds meal (i.e. 0% w/w, 20% w/w, 30% w/w and 40% w/w) for 180 days. The presence of transgenic DNA fragments (Cry1Ac, Cry2A and CP4 EPSPS) or plant endogenous gene (Sad1) was traced in those specific tissues and organs. The presence of ß-actin (ACTB) was also monitored as an internal control. Neither the transgenic fragments (459 bp of Cry1Ac gene, 167 bp of Cry2A gene and111 bp of CP4 EPSPS gene) nor cotton endogenous reference gene (155 bp of Sad1) could be detected in any of the DNA samples extracted from the rabbit's tissues in both control and transgenic groups. However, 155 bp fragment of the rabbit's reference gene (ACTB) was recovered in all the DNA samples extracted from rabbit tissues. The results obtained from this study revealed that both plant endogenous and transgenic DNA fragments have same fate in rabbit's tissues and were efficiently degraded in the gastrointestinal tract (GIT).

Nigerian surgical outcomes - Report of a 7-day prospective cohort study and external validation of the African surgical outcomes study surgical risk calculator.

BACKGROUND: Surgical outcomes study for individual nations remains important because of international differences in patterns of surgical disease. We aimed to contribute to data on post-operative complications, critical care admissions and mortality following elective surgery in Nigeria and also validate the African Surgical Outcomes Study (ASOS) surgical risk calculator in our adult patient cohort. MATERIALS AND METHODS: We conducted a 7-day, national prospective observational cohort study in consented consecutive patients undergoing elective surgery with a planned overnight hospital stay following elective surgery during a seven-day study period. The outcome measures were in-hospital postoperative complications, critical care admissions and in-hospital mortality censored at 30 days. Also, we identified variables which significantly contributed to higher ASOS surgical risk score. External validation was performed using area under the receiver operating characteristic curve (ROC) for discrimination assessment and Hosmer-Lemeshow test for calibration. RESULTS: A total of 1,425 patients from 79 hospitals participated in the study. Postoperative complications occurred in 264(18.5%, 95% CI 16.6-20.6), 20(7.6%) of whom were admitted into the ICU and 16(6.0%) did not survive. Total ICU admission was 57 (4%), with mortality rate of 23.5% following planned admission and overall in-hospital death was 22(1.5%, 95% CI 0.9-2.2). All prognostic factors in the ASOS risk calculator were significantly associated with higher ASOS score and the scoring system showed moderate discrimination (0⋅73, 95% CI 0.62-0.83). Hosmer-Lemeshow χ2 test revealed scale was well calibrated in the validation cohort. CONCLUSION: NiSOS validates the findings of ASOS and the ability of the ASOS surgical risk calculator to predict risk of developing severe postoperative complications and mortality. We identified failure-to-rescue as a problem in Nigeria. Furthermore, this study has provided policy makers with benchmarks that can be used to monitor programmes aimed at reducing the morbidity and mortality after elective surgery. We recommend the adoption of the ASOS surgical risk calculator as a tool for risk stratification preoperatively for elective surgery.

A modified protocol for rapid DNA isolation from cotton (Gossypium spp.).

Extraction of high-quality DNA from Gossypium (Cotton) species is notoriously difficult due to high contents of polysaccharides, quinones and polyphenols other secondary metabolites. Here, we describe a simple, rapid and modified procedure for high-quality DNA extraction from cotton, which is amenable for downstream analyses. In contrast to other CTAB methods, the described procedure is rapid, omits the use of liquid nitrogen, phenol, CsCl gradient ultracentrifugation, uses inexpensive and less hazardous reagents, and requires only ordinary laboratory equipment. The procedure employed the high concentration of NaCl and use of PVP-10 to rid the problems associated with polysaccharides and polyphenols, respectively. The average yield was approximately 10-15 µg of good quality DNA from 100 mg of tissue weight, which is adequate for projects, like genetic mapping or marker-assisted plant breeding. This protocol can be performed in as little as 3 h and may be adapted to high-throughput DNA isolation. •Buffers A and B were redesigned from Paterson et al. (1993) and Porebski et al. (1997), respectively.•Ribonuclease A was added before chloroform extraction.•A simple, rapid and inexpensive DNA extraction method is described.

Effect of dietary supplementation of recombinant Cry and Cp4 epsps proteins on haematological indices of growing rabbits.

Genetically modified (GM) crops expressing insect resistance and herbicide tolerance provide a novel approach for improved crop production but their advent at the same time presents serious challenges in terms of food safety. Although prevailing scientific proof has suggested that transgenic crops are analogous to their conventional counterparts, their use in human and animal diet gave rise to emotional public discussion. A number of studies had been conducted to evaluate the potential unintended effects of transgenic crops expressing single transgene, but very few studies for those with multiple transgenes. As the crops with single and multiple transgenes could impart different effects on non-target organisms, thus, risk evaluation of transgenic crops expressing more than one transgene is required to declare their biosafety. The present study was therefore designed to assess the effects of different levels of dietary transgenic cottonseed expressing recombinants proteins produced by Cry1Ac, Cry2A and Cp4epsps genes on haematological indices of growing rabbits. A total of 48 rabbits were assigned to four dietary treatments containing different levels of transgenic cottonseeds (i.e., 0% w/w, 20% w/w, 30% w/w and 40% w/w) with 0% w/w serving as control. Haematological parameters were measured at periodic intervals (0, 45, 90, 135 and 180) days. No significant (p > 0.05) dose-dependent effects were observed in most of the haematological parameters evaluated. Though, significant differences (p < 0.05) were recorded in the level of MCHC, MCH and HCT in some of experimental male and female rabbits, yet, they were not biologically significant, as all the differences were within the normal reference values. Our study suggested that feeding transgenic cottonseed of up to 40% could not adversely affect rabbit's haematological profile. However, further study needs to be conducted with different cotton genotypes expressing both single and polygenic traits before recommending the utilization of transgenic cottonseed in routine livestock feeding.

A case of rabies in a Kano brown doe.

This manuscript aims at creating awareness especially to livestock farmers and veterinarians/clinicians that rabies occurs in livestock species. Furthermore, clinicians should note that the viral load in dog bite cases by rabid dog can be reduced by vigorous washing and disinfection which may ultimately delay the clinical manifestation of rabies.

Nutritional Assessment of Dietary Bt and Cp4EPSPS Proteins on the Serum Biochemical Changes of Rabbits at Different Developmental Stages.

In recent years, the influence of genetically modified (GM) cotton expressing different types of Bt and EPSPS genes has been attested in term of reduced application of pesticides and insecticides coupled with improved cotton production. Although the cultivation of GM cotton has been authorized by the regulatory authorities of various countries in the world, based on the biosafety studies reported by most of the GM cotton producers, yet the information on its safe use are inadequate. In order to support the issues on food safety, it is therefore mandatory to conduct further safety assessment studies on GM cotton for each independent transgenic event on the basis of case assessment rule. In the present study, the effects of different doses of dietary GM cotton seed expressing Bt and EPSPS genes were studied on the level of serum biochemical in albino rabbits (Oryctolagus cuniculus). The rabbits were fed a diet containing different levels of GM cotton seeds (i.e., 20, 30, and 40% w/w) respectively mixed with standard diet for 180 days. During the course of the study, various serum enzymes, electrolytes, proteins, glucose and serum total cholesterol were examined at specific time intervals (0, 45, 90, 135, and 180) days. The results showed non-significant (P > 0.05) dose dependent effects in most of the evaluated serum biochemical parameters. Although, the results in some of the serum biochemistry were significantly different (P < 0.05) among the groups, however, they were not biologically significant, since all the differences were within the normal physiological range. These results thus, suggested that the GM cotton seed meal could be considered as safe as other conventional feed ingredients. The experimental evidence for the safe usage of GM cotton was highlighted in this study.

Molecular Approaches for High Throughput Detection and Quantification of Genetically Modified Crops: A Review.

As long as the genetically modified crops are gaining attention globally, their proper approval and commercialization need accurate and reliable diagnostic methods for the transgenic content. These diagnostic techniques are mainly divided into two major groups, i.e., identification of transgenic (1) DNA and (2) proteins from GMOs and their products. Conventional methods such as PCR (polymerase chain reaction) and enzyme-linked immunosorbent assay (ELISA) were routinely employed for DNA and protein based quantification respectively. Although, these Techniques (PCR and ELISA) are considered as significantly convenient and productive, but there is need for more advance technologies that allow for high throughput detection and the quantification of GM event as the production of more complex GMO is increasing day by day. Therefore, recent approaches like microarray, capillary gel electrophoresis, digital PCR and next generation sequencing are more promising due to their accuracy and precise detection of transgenic contents. The present article is a brief comparative study of all such detection techniques on the basis of their advent, feasibility, accuracy, and cost effectiveness. However, these emerging technologies have a lot to do with detection of a specific event, contamination of different events and determination of fusion as well as stacked gene protein are the critical issues to be addressed in future.

Effect of the Prevalence of HIV/AIDS and the Life Expectancy Rate on Economic Growth in SSA Countries: Difference GMM Approach.

The productivity of countries around the globe is adversely affected by the health-related problems of their labour force. This study examined the effect of the prevalence of human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) and life expectancy on the economic growth of 33 Sub-Saharan African (SSA) countries over a period of 11 years (2002-2012). The study employed a dynamic panel approach as opposed to the static traditional approach utilised in the literature. The dynamic approach became eminent because of the fact that HIV/AIDS is a dynamic variable as its prevalence today depends on the previous years. The result revealed that HIV/AIDS is negatively correlated with economic growth in the region, with a coefficient of 0.014, and significant at the 1% level. That is, a 10% increase in HIV/AIDS prevalence leads to a 0.14% decrease in the GDP of the region. Tackling HIV/AIDS is therefore imperative to the developing Sub-Saharan African region and all hands must be on deck to end the menace globally.