Exploring the genetic diversity of Eimeria acervulina: A polymerase chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) approach.

Eimeria, protozoan parasites that can cause the disease coccidiosis, pose a persistent challenge to poultry production and welfare. Control is commonly achieved using good husbandry supplemented with routine chemoprophylaxis and/or live parasite vaccination, although widespread drug resistance and challenges to vaccine supply or cost can prove limiting. Extensive effort has been applied to develop subunit anticoccidial vaccines as scalable, cost-effective alternatives, but translation to the field will require a robust understanding of parasite diversity. Using a new Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) panel we begin to describe the genetic diversity of Eimeria acervulina populations in Africa and Europe. PCR-RFLP genotyping E. acervulina populations sampled from commercial broiler and layer chickens reared in Nigeria or the United Kingdom (UK) and Republic of Ireland (RoI) revealed comparable levels of haplotype diversity, in direct contrast to previous descriptions from the close relative E. tenella. Here, 25 distinct PCR-RFLP haplotypes were detected from a panel of 42 E. acervulina samples, including 0.7 and 0.5 haplotypes per sample in Nigeria (n = 20) and the UK/RoI (n = 14), respectively. All but six haplotypes were found to be country-specific. The PCR-RFLP markers immune mapped protein 1 (IMP1) and heat shock protein 90 (HSP90) were most informative for Nigerian E. acervulina, while microneme protein 3 (MIC3) and HSP90 were most informative in UK/RoI populations. High haplotype diversity within E. acervulina populations may indicate frequent genetic exchange and potential for rapid dissemination of genetic material associated with escape from selective barriers such as anticoccidial drugs and future subunit vaccines.

Bioaccumulation of organochlorine pesticides in the parasite Cosmocerca sp. (Nematoda: Cosmocercidae) and the amphibian host Amietophrynus regularis (Reuss, 1833) within Lagos metropolis, Nigeria.

BACKGROUND: Amphibian species are being threatened worldwide and chemical pollution is one of the leading causes of this decline. The use of agrochemicals such as organochlorine pesticides (OCPs) among the several health and ecological challenges it causes, the sharp amphibian population decline is most pressing.Toad specimens Amietophrynus regularis were sampled from three (3) selected areas; each comprising of natural habitat and dumpsites within Lagos metropolis. METHODS: The congeners of organochlorine pesticides were tested in the liver, intestine, and parasite (Cosmocerca sp.) of the toads and soil samples from the respective locations using gas chromatography-mass spectrometer (GC-MS). Histopathological analyses were conducted on the intestines and liver of the toads using hematoxylin and eosin (H&E) stain and then examined under the binocular dissecting microscope. RESULTS: The concentration of aldrin in the intestine of A. regularis sampled at the dumpsites was higher than the concentrations in the intestines of A. regularis in the natural habitat. The concentrations of dieldrin in the uninfected A. regularis at both dumpsite and natural habitat were higher than the concentrations in the infected A. regularis at both environments. This indicated that the parasite Cosmocerca sp. may have played a depurative role in sequestering the concentration of dieldrin in the toads irrespective of the location. The parasites exhibited marked sequestration capacity characterized by the notably high total bioaccumulation rate both in the liver and the intestine at the dumpsite. The stunted villi being the common histological alteration in the infected and uninfected toads at the dumpsite but missing in the uninfected counterparts at the natural habitat may be attributed to the differences in the background concentration of the OCP congeners. CONCLUSIONS: The parasite- Cosmocerca sp. has been shown to be a potential tool in the biomonitoring of these OCP congeners which persists in the environment. Continuous research on these congeners is a searchlight to checkmate the environment to see how compliant industries and the consumers are in terms of regulation of these chemicals.

Studies on heavy metals and fish health indicators in Malapterurus electricus from Lekki Lagoon, Lagos, Nigeria.

The aquatic ecosystem is constantly being disturbed by rising levels of different classes of pollutants of human origin in the form of urban, agricultural and industrial discharges. In this study, the health of fish Malapterurus electricus was examined, to serve as a reflection of the impact of anthropogenic disturbances in the Lekki lagoon. Eighty six samples of the fish were analysed for parasitic infections, heavy metals, microorganisms in their internal and external body parts while the proximate composition and fish condition factor were also determined using conventional methods. One parasite species, a cestode Electrotaenia malapteruri was found to infect the fish. Total parasite load was eighty five with a total prevalence of infection of 36%. Elements detected in sediment were of the order of Al>Fe>Mn>Pb>Cr>Zn>Cd>Ba>Cu>Ni while in water, it is Mn>Fe>Zn>Ba>Cd>Cr>Al>Cu>Ni>Pb. Metals analysed in the fish tissues were generally low and below regulatory limits. In the proximate analysis, moisture content has a value of 80.7%, while ash content had a value of 1.26%. Eight bacterial and two fungal species were isolated from the fish. The condition factor of fish varied between 1 and 2. The study provides valuable information for monitoring and management of heavy metal pollution in the aquatic ecosystem.

Nilonema gymnarchi (Nematoda: Philometridae) and trace metals in Gymnarchus niloticus of Epe lagoon in Lagos State, Nigeria.

The presence of trace metals in the sediment, water, and biota of the Epe lagoon has been recently linked to oil exploration and municipal perturbations around the lagoon. The study was aimed at assessing the concentrations and associated health risks of Fe, Zn, Cu, Ni, Pb, Cd, Cr, Mn, Co and V in the water, sediment, and Gymnarchus niloticus of Epe lagoon and to evaluate the role of the enteric parasite Nilonema gymnarchi in bioaccumulation of the metals in the fish. The temperature, pH, redox potential, conductivity, turbidity, dissolved oxygen (DO), total dissolved solids (TDS), and salinity were determined in-situ using a handheld multi-parameter probe (Horiba Water Checker Model U-10). The concentrations of Fe, Zn, Cu, Ni, Pb, Cd, Cr, Mn, Co, and V were determined in the surface water, bottom sediment, Gymnarchus niloticus, and its enteric parasites, Nilonema gymnarchi in Epe lagoon using the Flame Atomic Absorption Spectrometer (Philips model PU 9100). The bioaccumulation factors and target hazard quotients of the trace metals in the infected and uninfected fish were estimated and compared. The intestinal tissue sections of the infected and uninfected fish were examined using a binocular dissecting microscope (American Optical Corporation, Model 570) using hematoxylin and eosin (H&E) stain. Biochemical markers such as reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), and lipid peroxidation (MDA) were determined in the liver of the infected and uninfected fish. The SOD level was higher in the uninfected fish than the infected ones supports the indications deduced from the bioaccumulation analysis. Strong positive correlations between SOD and most of the metals- Fe (0.916), Zn (0.919), Cu (0.896), and Ni (0.917) suggests that the metals may have inflicted more toxicity in the uninfected. The histopathological comparisons made between the uninfected and infected fish showed consistency with the outcomes of other comparisons made in this study. These evidence were marked by tissue alterations in the infected fish ranging from no observed changes to mild alterations, while the uninfected exhibited more severe tissue injuries such as hemorrhagic lesions, severe vascular congestion, edema, the increased connective tissue of the submucosa, and vascular congestion. The condition factors of the infected (0.252) and uninfected (0.268) fish indicated slenderness and unfitness possibly due to environmental stressors such as trace metals. The parasitized fish showing better-coping potentials than the uninfected, coupled with the significant bioaccumulation interferences exhibited by the parasite Nilonema gymnarchi is an indication that the parasites may be a good metal sequestration agent for the fish and can be used to forestall the significant health hazard quotient posed by the current level of iron and the synergy of all metals analyzed in the lagoon.

Heavy metals, parasitologic and oxidative stress biomarker investigations in Heterotis niloticus from Lekki Lagoon, Lagos, Nigeria.

Heavy metal toxicity in aquatic life as a result of human activities poses a grave health threat to water quality, aquatic and human life. Parasites may serve as indicators of heavy metal pollution. This research investigated the health status of the fish Heterotis niloticus viz-a-viz quality of the water and sediments in Lekki lagoon, parasitic infection, presence of heavy metals and oxidative stress response in the liver and intestine of the fish. Parasites recovered were also analyzed for the extent of bioaccumulation of heavy metals. The metals in water, sediments, parasites, and fish were analyzed using Atomic Absorption Spectrometry. Heavy metal concentrations in the surface water were generally below regulatory limits of World Health Organization. Sediment had high levels of aluminium (124.78 mg/kg) and iron (327.41 mg/kg); other heavy metals were below regulatory limits. Tenuisentis niloticus, an acanthocephalan, was the only parasite recovered. Seventy (70) out of 100 fish sampled were infected with the parasite. T. niloticus bioaccumulated Cd, Ni, and Pb between 65 to 100 times more than the liver and 12 to 200 times more than the intestine. Other metals bioaccumulated from the host tissues by the parasite had the magnitude between 1 to 12 times as the liver and 1 to 30 times as the intestine. There were significant differences in the activities of antioxidant enzymes between the parasitized and non-parasitized fishes. Fish tissues also showed histological alterations, ranging from mild infiltration of inflammatory cells to moderate inflammation and haemorrhagic lesions. Human activities that introduce stressors into the lagoon should be controlled.

Impacts of trace metals on African common toad, Amietophrynus regularis (Reuss, 1833) and depuration effects of the toad's enteric parasite, Amplicaecum africanum (Taylor, 1924) sampled within Lagos metropolis, Nigeria.

The study aimed at assessing the depuration potentials of endoparasite, Amplicaecum africanum on trace metals in its toad host, Amietophrynus regularis at sites of significant anthropogenic perturbations within the Lagos metropolis, in Nigeria. A total of 120 toads of both sexes, alongside 45 soil samples were collected from each of three (3) stations labeled Dumpsite, Lagoon front and Highrise, using hand nets and by hand-picking between February and October, 2018. The intestinal tissues sections of the toads were examined using a binocular dissecting microscope (American Optical Corporation, Model 570) and hematoxylin and eosin (H&E) stain. Oxidative stress in toad intestine was assessed by estimating the levels of glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT), and lipid peroxidation (MDA). Trace metals in the water, soil, toad liver, intestine and parasite, Amplicaecum africanum were tested using Atomic Absorption Spectrophotometry (Philips model PU 9100). Lead (Pb), copper (Cu), nickel (Ni), cadmium (Cd), and chromium (Cr) were detected in the toads, with the infected toads having lower concentrations of most trace metals than the uninfected toads, irrespective of the locations and sex. Strong negative correlations between parasitological indices and concentrations of trace metals in the toads suggest that the parasites might have taken up significant amounts of trace metals from the host. The study demonstrated the potentials of parasite, A. africanum to depurate trace metal burden in Amietophrynus regularis. When the dominant factor impacting the toad is the parasitic infection, parasite intensity determines the trade-off between parasitological harm and depuration benefit to the host. Hence, under controlled conditions, parasites may serve as bioremediation agent in the event of pollution. Depuration potential of A. africanum in the study was supported by the mild tissue alterations observed in the intestine of infected toads, compared to the uninfected counterparts, which exhibited severe glandular hyperplasia, increased connective tissue, and severely stunted villi. Consistently lower activities of biochemical biomarkers which characterize the uninfected toads compared to the infected, irrespective of the sex and stations, further corroborate drawn inferences.

Bioaccumulation of BTEX and PAHs in Heterotis niloticus (Actinopterygii) from the Epe Lagoon, Lagos, Nigeria.

Toxicity studies in the aquatic ecosystem have shown that petrol and its product have adverse effects on aquatic biota. They are able to easily absorb these toxic substances into their bodies from sediment, water or even food items that are contaminated, thus impacting the food chain. In this study, water, sediment and fish (Heterotis niloticus) from the Epe Lagoon were investigated for the presence of BTEX (benzene, toluene, ethylbenzene and the three xylene isomers m, o and p-xylenes), and PAHs using GC-MS. Total concentration of BTEX in sediment and water was respectively 32.37 ± 1.07 µg/g, 49.86 ± 7.30 µg/L; while total concentration of BTEX in fish liver and intestine was 33.75 ± 10.09 and 40.16 ± 9.64 µg/g respectively. Benzene, 1,2-Dichlorobenzene and 1,3-Dichlorobenzene components of BTEX were not detected in both sediment and water. Total concentration of PAH in sediment and water was 7.46 ± 1.73 µg/g and 19.33 ± 1.31 µg/L respectively. Total PAHs concentration in liver and fish intestine was 141.23 ± 5.89 and 173.34 ± 4.677 µg/g respectively. Majority of the PAHs congeners were found to be higher than the acceptable limits. Findings from this study shows the need for continuous monitoring of our natural waters as the present situation portends a potential concern for ecological risk.

Infectivity of Simulium damnosum s.l. and therapeutic coverage of ivermectin distribution 10 years post treatment around Owena Dam, Ondo state, Nigeria

ABSTRACT Background: Studies related to infectivity status of insect vectors are seen as necessities in understanding the epidemiology of vector-borne diseases and planning effective control measures. This study assessed the infectivity ofSimulium damnosum s.l. around Owena River as well as evaluated therapeutic coverage of Ivermectin distribution in the area. Method: Human landing sampling method was used to collect adult flies on human attractants from 07:00 to 18:00 for two consecutive days a month for three months (July 2016 - September 2016). Parity assessment was conducted to determine the age of fly populations. Parous flies were further dissected to detect the presence or absence ofOnchocerca larvae. Biting rates and transmission potentials were calculated using standard methods. A quantitative survey was carried out to determine the therapeutic coverage and compliance to ivermectin treatment for the control of Onchocerciasis in the study area using standard household coverage questionnaires. Results: A total of 914 adult female flies were collected during the study period. The daily biting rate (DBR) varied from 146 fly per man day (FMD) in July to 162.5 FMD in August. The monthly biting rate (MBR) was lowest in September (2170 bites per man per month) but highest in August (3358.3 bites per man per month). MBD ranged from 13.23 fly per man hour (FMH) in July to 14.77 FMH in August. The results indicated that the majority of the flies collected at the sampling points were nulliparous [685 (74.95%)] while others were parous [229 (25.05%)]. The biting activity of the flies showed a marked decrease in population in August compared to July which later increased in September. Infection rates varied from 2 (0.7%) in July to 7 (2.2%) in August while the infectivity rate during the study ranged from zero (July and September) to 3 (1.0%) in August. Conclusion: Despite the years of treatment of onchocerciasis in Owena community, there were still some infective flies capable of transmitting O. volvolus. This could be due to the low rate of therapeutic coverage as a result of non-compliance in the community for various reasons earlier stated.

Infectivity of Simulium damnosum s.l. and therapeutic coverage of ivermectin distribution 10 years post treatment around Owena Dam, Ondo state, Nigeria.

BACKGROUND: Studies related to infectivity status of insect vectors are seen as necessities in understanding the epidemiology of vector-borne diseases and planning effective control measures. This study assessed the infectivity ofSimulium damnosum s.l. around Owena River as well as evaluated therapeutic coverage of Ivermectin distribution in the area. METHOD: Human landing sampling method was used to collect adult flies on human attractants from 07:00 to 18:00 for two consecutive days a month for three months (July 2016 - September 2016). Parity assessment was conducted to determine the age of fly populations. Parous flies were further dissected to detect the presence or absence ofOnchocerca larvae. Biting rates and transmission potentials were calculated using standard methods. A quantitative survey was carried out to determine the therapeutic coverage and compliance to ivermectin treatment for the control of Onchocerciasis in the study area using standard household coverage questionnaires. RESULTS: A total of 914 adult female flies were collected during the study period. The daily biting rate (DBR) varied from 146 fly per man day (FMD) in July to 162.5 FMD in August. The monthly biting rate (MBR) was lowest in September (2170 bites per man per month) but highest in August (3358.3 bites per man per month). MBD ranged from 13.23 fly per man hour (FMH) in July to 14.77 FMH in August. The results indicated that the majority of the flies collected at the sampling points were nulliparous [685 (74.95%)] while others were parous [229 (25.05%)]. The biting activity of the flies showed a marked decrease in population in August compared to July which later increased in September. Infection rates varied from 2 (0.7%) in July to 7 (2.2%) in August while the infectivity rate during the study ranged from zero (July and September) to 3 (1.0%) in August. CONCLUSION: Despite the years of treatment of onchocerciasis in Owena community, there were still some infective flies capable of transmitting O. volvolus. This could be due to the low rate of therapeutic coverage as a result of non-compliance in the community for various reasons earlier stated.

Toxicological effects of prolonged and intense use of mosquito coil emission in rats and its implications on malaria control.

Mosquito coil is a vector control option used to prevent malaria in low income counties, while some studies have addressed this issue, additional reseach is required to increase knowledge on the adverse health effects caused by the prolonged use of coils. In this study we investigated the toxicological effects of fumes from two locally manufactured mosquito coil insecticides (with pyrethroids: transfluthrin and d-allethrin as active ingredients) on male albino rats. For this, we recorded the haematological and biochemical indices, and made histopathology and mutagenicity evaluations in rats exposed to mosquito fumes during 2, 4, 8, 12 and 16 week periods. Haematological determination was performed using automated hematology analyzer to determine White Blood Cell (WBC), Packed Cell Volume (PCV), Red Blood Cell (RBC) and Platelet (PLT) counts, while biochemical evaluations were determined using available commercial kits. Gross histopathological changes were studied for the kidney, liver and lungs in sacrificed rats. The rat sperm head abnormalities assessment was used to evaluate mutagenicity. Mosquito coil fumes produced significant increase (P < 0.05) in the levels of total protein, total albumin and bilirubin, when animals were exposed from two weeks to 16 weeks with transfluthrin. Similarly, elevation in the activities of aspartate amino transferase, alanine amino transferase and alanine phosphatase, increased significantly in both insecticides. Increase in WBC, RBC and PCV were recorded for all the exposure periods, however PLT count showed no significant increase (P > 0.05). Mutagenicity assessment revealed sperm abnormality was statistically significant (P < 0.05) compared with the control at 8, 12 and 16 weeks post exposure to transfluthrin. Histological studies revealed severe lung damage evidenced by interstitial accumulations, pulmonary oedema and emphysema in exposed rats. Intracellular accumulations and severe sinusoidal congestion of liver cells were observed from 12 weeks exposure, indicating liver damage. Our studies indicate that mosquito coil fumes do initiate gradual damage to the host. These pathological effects must be taken into consideration by the malaria control program, particularly when regulating their long term and indoor usage.

Toxicological effects of prolonged and intense use of mosquito coil emission in rats and its implications on malaria control / Efectos toxicológicos del uso prolongado e intenso de emisiones de espirales contra mosquitos en ratas y sus implicaciones sobre el control de la malaria

Efectos toxicológicos del uso prolongado e intenso de emisiones de espirales contra mosquitos en ratas y sus implicaciones sobre el control de la malaria. Mosquito coil is a vector control option used to prevent malaria in low income counties, while some studies have addressed this issue, additional reseach is required to increase knowledge on the adverse health effects caused by the prolonged use of coils. In this study we investigated the toxicological effects of fumes from two locally manufactured mosquito coil insecticides (with pyrethroids: transfluthrin and d-allethrin as active ingredients) on male albino rats. For this, we recorded the haematological and biochemical indices, and made histopathology and mutagenicity evaluations in rats exposed to mosquito fumes during 2, 4, 8, 12 and 16 week periods. Haematological determination was performed using automated hematology analyzer to determine White Blood Cell (WBC), Packed Cell Volume (PCV), Red Blood Cell (RBC) and Platelet (PLT) counts, while biochemical evaluations were determined using available commercial kits. Gross histopathological changes were studied for the kidney, liver and lungs in sacrificed rats. The rat sperm head abnormalities assessment was used to evaluate mutagenicity. Mosquito coil fumes produced significant increase (P<0.05) in the levels of total protein, total albumin and bilirubin, when animals were exposed from two weeks to 16 weeks with transfluthrin. Similarly, elevation in the activities of aspartate amino transferase, alanine amino transferase and alanine phosphatase, increased significantly in both insecticides. Increase in WBC, RBC and PCV were recorded for all the exposure periods, however PLT count showed no significant increase (P>0.05). Mutagenicity assessment revealed sperm abnormality was statistically significant (P<0.05) compared with the control at 8, 12 and 16 weeks post exposure to transfluthrin. Histological studies revealed severe lung damage evidenced by interstitial accumulations, pulmonary oedema and emphysema in exposed rats. Intracellular accumulations and severe sinusoidal congestion of liver cells were observed from 12 weeks exposure, indicating liver damage. Our studies indicate that mosquito coil fumes do initiate gradual damage to the host. These pathological effects must be taken into consideration by the malaria control program, particularly when regulating their long term and indoor usage.

Las espirales contra los mosquitos se utilizan en los países de bajos ingresos como una opción para prevenir la malaria controlando el vector de esta enfermedad. A pesar de que algunos estudios han abordado este tema, se requiere más investigación para incrementar el conocimiento sobre los efectos adversos en la salud, causados por el uso prolongado de las espirales. En este estudio se investigaron los efectos toxicológicos de los gases de las espirales a partir de dos insecticidas fabricados en el país (con piretroides: transflutrina y d-aletrina como ingredientes activos) en machos de ratas albinas. Para esto, se registraron los índices hematológicos y bioquímicos, y se hicieron evaluaciones histopatológicas y de mutagenicidad en ratas expuestas a los gases de las espirales durante períodos de 2, 4, 8, 12 y 16 semanas. La determinación hematológica se realizó mediante un analizador de hematología automatizado para determinar el conteo de los Glóbulos Blancos (WBC), el Hematocrito (PCV), Glóbulos Rojos (RBC) y las Plaquetas (PLT), mientras que las evaluaciones bioquímicas se determinaron utilizando kits comerciales disponibles. Los cambios histopatológicos fuertes se estudiaron en el riñón, el hígado y los pulmones de ratas sacrificadas. Las anormalidades en la cabeza de los espermatozoides de las ratas se utilizaron para evaluar la mutagenicidad. El humo de las espirales contra los mosquitos producen un aumento significativo (p<0.05) en los niveles de proteína total, albúmina total y bilirrubina, cuando los animales fueron expuestos de dos semanas a 16 semanas con transflutrina. Del mismo modo, la elevación en las actividades de aspartato amino transferasa, alanina amino transferasa y alanina fosfatasa, aumentó significativamente con ambos insecticidas. Se registro un aumento en los leucocitos, eritrocitos y el hematocrito para todos los períodos de exposición, sin embargo el recuento de las plaquetas no mostró un aumento significativo (p>0.05). Las pruebas de mutagenicidad revelaron que las anormalidades en el esperma de las ratas fue estadísticamente significativa (p>0.05) al comparar el control a las 8, 12 y 16 semanas post exposición a la transflutrina. Los estudios histológicos revelaron una serie de daños pulmonares graves en las ratas expuestas al humo de la espiral, evidenciados por la acumulación intersticial, edema pulmonar y enfisema. Las acumulaciones intracelulares y la congestión sinusoidal severa de las células del hígado se observaron a partir de las 12 semanas de exposición, lo que indica daño hepático. Nuestros estudios indican que los vapores de las espirales contra mosquitos inician el daño gradual al huésped. Estos efectos patológicos deben ser tomados en cuenta por el programa de control de la malaria, particularmente a la hora de regular su uso a largo plazo y bajo techo.