Long term integrated biomarker responses in freshwater African catfish Clarias gariepinus exposed to a new brand of herbicide fluazifop-p-butyl.

The study investigated the sublethal effects of Fluazifop-p-butyl (FPB) on the haematological, biochemical and oxidative stress changes in Clarias gariepinus. Juvenile C. gariepinus were exposed to FPB concentrations of 1.80, 3.50, and 7.10 mg/l corresponding to 5, 10 and 20% of 96 h LC50 value of FPB respectively for 21 days and allowed to recover for 7 days. The blood, liver and gills were removed and analyzed. Fish exposed to different concentrations of FPB showed significant decline in the values the pack cell volume, hemoglobin and red blood cells but the white blood cell values increased. The neutrophil values increased while the lymphocyte declined but the monocytes, basophil and eosinophil values remain unchanged. The liver and gill aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase significantly increased compared to the control. There were mixed trends in the values of glucose but total protein was reduced in both tissues of the fish. There was significant decline in superoxide dismutase, catalase and glutathione peroxidase while malondialdehyde, total glutathione, and glutathione reductase increased in both liver and gill of the exposed fish. Following the 7-day withdrawal, most of the observed parameters returned to normal values. The present study revealed that FPB is toxic to C. gariepinus juveniles and prolonged exposure could result to major health risks to aquatic organisms, hence, they should be carefully used.

Sources, pollution, and ecological risk assessment of polycyclic aromatic hydrocarbons (PAHs) in Porto-Novo Lagoon, Benin Republic.

The Porto-Novo Lagoon is influenced by agricultural discharges and human activities. In order to evaluate the impact of wastes and human activities on Porto-Novo Lagoon, the sources and ecological risks of sixteen polycyclic aromatic hydrocarbons (PAHs) were assessed. The physicochemical and biological parameters of the water were also determined. The result showed that between the sampling sites, the mean concentration of dissolved oxygen ranged from 4.8 ± 0.5 to 5.1 ± 0.2 mg/L; biochemical oxygen demand varied from 12.6 ± 2.0 to 77.9 ± 81.9 mg/L; biological oxygen demand ranged from 2.8 ± 2.6 to 5.6 ± 0.9 mg/L; total phosphorus varied between 4.7 ± 2.7 and 15.3 ± 9.5 mg/L; total dissolved solids ranged from 183.0 ± 115.8 to 337.5 ± 413.3 mg/L, and Escherichia coli varied from 495.0 ± 542.9 to 1920.0 ± 2676.5 UFC/100 mL. Water parameter values obtained were not within World Health Organization (WHO)-recommended limits except pH and TDS. Total PAHs (∑PAHs) concentration varied from 38.8 to 123.9 mg/L. The mean ∑PAH concentration was 83.2 ± 20.3 mg/L. Benzo[b]fluoranthene, benzo[g,h,i]perylene, and benzo[k]fluoranthene were the most dominant PAHs and contributed to 55.9%, 15.3%, and 4.5% of the ∑PAHs concentration, respectively. Douane-Tokpa and Djassin recorded the lowest and highest concentrations. PAHs of four to six rings were the most abundant across the sampling sites. Naphthalene showed the lowest risk in the lagoon. Acenaphthene showed low risk at Djassin, while Indeno(1,2,3 cd)pyrene showed low risk at Benin Industry Body Fat. Except for those that were not detected, all the PAHs at individual or complex mixture levels showed high risk at all the sites. The highest total concentration was recorded in Djassin followed by Beaurivage. The high level of PAHs pollution was attributed to both human and goods traffic, runoff, and the complex hotels close to the lagoon. Molecular diagnostic ratios and principal component analysis suggest that the target hydrocarbons were from both petrogenic and pyrogenic sources with predomination of vehicular emission and coal/woods combustion. ∑LWM/HWM confirmed also the predominance of pyrolytic sources of PAHs in Porto-Novo Lagoon. The predominance of the vehicular emission may be due to the position of the complex Porto-Novo Lagoon-Nokoué Lake which is between the two big cities of the country.

Biomorphometric and hematobiochemical alterations in the juvenile african catfish Clarias gariepinus exposed to propranolol.

Propranolol (PRO) is a beta-blocker drug used for the treatment of anxiety, chest pain, migraine and tremors. The present study investigated whether sublethal concentrations of PRO have effects on the body condition, biochemistry, and hematology of Clarias gariepinus juveniles. The 96-h median lethal concentration (LC50) of the drug, which was established through an acute toxicity study, was 9.48 mg/L. Based on these values, the fish were exposed for 21 days to the control and sublethal concentrations of 1.90, 0.95, and 0.63 mg/L, which are equivalent to the 1/5th, 1/10th, and 1/20th of the LC50 of PRO, respectively. After 21 days of exposure, the fish were removed from the toxicant and kept in toxicant-free water for 7 days to recover. The standard length and body weight of each fish were measured after each exposure period. The condition factor (CF) and hepatosomatic index (HSI) were not significantly affected by the drug. The red blood cell (RBC) count, hemoglobin (Hb) count and packed cell volume (PCV) decreased from day 7 to 21 at the tested concentrations, while the white blood cell (WBC) count significantly increased. There were alterations in the mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and mean corpuscular hemoglobin (MCH) in the exposed groups compared to those in the control group. When neutrophil counts increased, the lymphocyte counts decreased, but the monocyte, basophil cell and eosinophil cell counts were not affected. Among the liver enzymes, only aspartate aminotransferase was significantly stimulated in the groups that were exposed to the drug. The protein and glucose levels of fish exposed to the drug decreased. Most of the studied parameters returned to their original values after the 7-day recovery period. The information provided in the current study will be helpful in the monitoring of PRO contamination in aquatic environments.

Hematological, biochemical, and biometric changes in Clarias gariepinus exposed to antipsychotic drug chlorpromazine.

Chlorpromazine (CPZ) is a neuroleptic and antipsychotic medication for individuals suffering from schizophrenia and other medical conditions. This study investigated the effects of CPZ on the hematological, biochemical, and biometric characteristics in juvenile Clarias gariepinus. The fish were exposed to 0.53, 1.06, and 2.11 mgL-1 CPZ for 15 days after which they were withdrawn from the toxicant and allowed to recover for 5 days. Blood were sampled from the fish on days 1, 5, 10, 15, and during the 5-day recovery for hematological and biochemical analysis, and thereafter, the fish were sacrificed for the morphometric analysis. While the values of the white blood cells significantly increased in the exposed fish, the hemoglobin, red blood cells, and packed cell volume decreased. Compared with the control, there were no significant differences in the values of the blood derivatives in the exposed fish. The values of protein and glucose reduced, but those of aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase were significantly elevated. Though there was no significant difference in the condition factor, a significant increase in hepatosomatic index occurred on day 15 at 5.28 mg/L CPZ. After the 5-day withdrawal from the drug, most of the studied parameters returned to the control values. The present study indicated that CPZ is toxic to fish and should be used with utmost care to guard against toxicological effect on non-target organisms.

Population structure and evolutionary history of the greater cane rat (Thryonomys swinderianus) from the Guinean Forests of West Africa.

Grasscutter (Thryonomys swinderianus) is a large-body old world rodent found in sub-Saharan Africa. The body size and the unique taste of the meat of this major crop pest have made it a target of intense hunting and a potential consideration as a micro-livestock. However, there is insufficient knowledge on the genetic diversity of its populations across African Guinean forests. Herein, we investigated the genetic diversity, population structures and evolutionary history of seven Nigerian wild grasscutter populations together with individuals from Cameroon, Republic of Benin, and Ghana, using five mitochondrial fragments, including D-loop and cytochrome b (CYTB). D-loop haplotype diversity ranged from 0.571 (± 0.149) in Republic of Benin to 0.921 (± 0.013) in Ghana. Within Nigeria, the haplotype diversity ranged from 0.659 (± 0.059) in Cross River to 0.837 (± 0.075) in Ondo subpopulation. The fixation index (FST), haplotype frequency distribution and analysis of molecular variance revealed varying levels of population structures across populations. No significant signature of population contraction was detected in the grasscutter populations. Evolutionary analyses of CYTB suggests that South African population might have diverged from other populations about 6.1 (2.6-10.18, 95% CI) MYA. Taken together, this study reveals the population status and evolutionary history of grasscutter populations in the region.

Morphometric, hematological and oxidative stress changes in Clarias gariepinus following sub-chronic exposure to tramadol.

Tramadol is among the most famous analgesic drugs used for the management, treatment and relief of moderate to severe pain conditions. The present study investigated the effects of tramadol on the behavior, mortality, morphometric, hematology and oxidative stress parameters of C. gariepinus juveniles. The 96 h LC50 value of tramadol determined by probit analysis was 88.76 mg/L. Based on this value, fish were exposed to sublethal concentrations of 4.44, 8.88, 17.75 mg/L tramadol and 0.0 mg/L (control) for the period of 15 days and allowed to recover for 5 days. Fish exposed to tramadol showed some abnormal behavioral responses and mortality increased with increase in the exposure duration and concentrations except for the control. There were variations in hepatosomatic index (HSI) and condition factor (CF) in fish exposed to tramadol. Exposure of C. gariepinus to tramadol elicited reduction in the values of white blood cell (WBC), red blood cell (RBC), hemoglobin (Hb), packed cell volume (PCV) and mean corpuscular volume (MCV) while the values of mean corpuscular hemoglobin (MCH) and the mean corpuscular hemoglobin concentration (MCHC) increased. The values of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR), reduced glutathione (GSH) and lipid peroxidation (LPO) increased significantly in the exposed fish compared with the control. The values of glutathione peroxidase (GPx) however decreased. The results of the present study demonstrate that tramadol is toxic to fish and its use should be monitored in the aquatic environment.

Seasonal variation and diversity of rotifers in Ehoma lake, Nigeria.

The zooplankton and physicochemistry of Ehoma lake were studied from March 2005 to August 2006 in order to evaluate seasonal trend in species composition and abundance in relation to water quality. Forty two (42) species of rotifers dominated by Keratella tropica tropica were encountered. Species richness declined by nearly 50% from dry season values during the rainy season. Species richness was higher in the littoral zone during the dry season, the reverse was the case during the rainy season. Species abundance followed similar trend as species richness. Abundance was closely associated with dissolved oxygen, biological oxygen demand and width of the lake and these in turn varied significantly between seasons. Shannon-Weaver diversity varied from 2.2 to 3.8 bit ind(-1) and significantly between seasons (p<0.05). Anuraeopsis navicula navicula and Euchlanis dilatata dilatata Ehrenbreg were the only species gained during the flood periods.

Morphological, haematological and biochemical changes in African catfish Clarias gariepinus (Burchell 1822) juveniles exposed to clotrimazole.

Clotrimazole (CLO) is an imidazole fungicide used in human and veterinary medicine for treating fungal infection. This study evaluated the changes in morphological, haematological and biochemical indices in Clarias gariepinus juveniles exposed to CLO. After the acute exposure, the 96 h LC50 value of CLO determined by probit analysis was 38.79 mgl-1. Based on this value, fish were exposed to sublethal concentrations of 7.76, 3.89, 1.94 and 0.00 mgl-1 (control) of CLO for 21 days and were allowed to recover for 7 days. The result revealed no significant effect on the hepatosomatic index and condition factor of the exposed fish. There were concentration and time-dependent significant decreases in red blood cell (RBC), haemoglobin (Hb), packed cell volume (PCV) and mean corpuscular volume (MCV) with significant increase in the white blood cell (WBC), mean corpuscular haemoglobin concentration (MCHC), and mean corpuscular haemoglobin (MCH) in the exposed group when compared with the control. A mixed trend was observed in the levels of neutrophils, lymphocytes, monocytes and eosinophils. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and glucose values significantly increased, while protein levels were reduced (p < 0.05) throughout the 21-day exposure and the 7-day recovery period. The present research indicated that CLO may have potential toxic effect on non-target organisms especially fish and, therefore, should be monitored in the aquatic ecosystem.

DNA Barcoding Silver Butter Catfish (Schilbe intermedius) Reveals Patterns of Mitochondrial Genetic Diversity Across African River Systems.

The silver butter catfish (Schilbe intermedius) is widely distributed across African river systems. To date, information on its mitochondrial genetic diversity, population structure, and historical demography are not well-established. Herein, we combined newly generated mitochondrial cytochrome c oxidase (COI) subunit I gene sequences with previously published COI sequences in the global databases to reconstruct its phylogeography, population genetic structure, and historical demography. Results from the mtDNA phylogeography and species delimitation tests (Cluster algorithm - Species Identifier, Automatic Barcode Gap Discovery and Poison Tree Process model) revealed that S. intermedius comprises at least seven geographically defined matrilines. Although the overall haplotype diversity of S. intermedius was high (h = 0.90), results showed that East (Kenya) and West (Nigeria) African populations had low levels of haplotype diversity (h = ~0.40). In addition, population genetic polymorphism and historical demographics showed that S. intermedius populations in both East and West Africa underwent severe contractions as a result of biogeographic influences. The patterns of genetic diversity and population structure were consistent with adaptive responses to historical biogeographic factors and contemporary environmental variations across African river systems. This is suggestive of the influence of historical biogeographic factors and climatic conditions on population divergence of S. intermedius across African river systems. Given our discovery of previously underappreciated diversity within S. intermedius, we recommend that this species be considered for increased conservation and management.

DNA barcoding of economically important freshwater fish species from north-central Nigeria uncovers cryptic diversity.

This study examines the utility of morphology and DNA barcoding in species identification of freshwater fishes from north-central Nigeria. We compared molecular data (mitochondrial cytochrome c oxidase subunit I (COI) sequences) of 136 de novo samples from 53 morphologically identified species alongside others in GenBank and BOLD databases. Using DNA sequence similarity-based (≥97% cutoff) identification technique, 50 (94.30%) and 24 (45.30%) species were identified to species level using GenBank and BOLD databases, respectively. Furthermore, we identified cases of taxonomic problems in 26 (49.00%) morphologically identified species. There were also four (7.10%) cases of mismatch in DNA barcoding in which our query sequence in GenBank and BOLD showed a sequence match with different species names. Using DNA barcode reference data, we also identified four unknown fish samples collected from fishermen to species level. Our Neighbor-joining (NJ) tree analysis recovers several intraspecific species clusters with strong bootstrap support (≥95%). Analysis uncovers two well-supported lineages within Schilbe intermedius. The Bayesian phylogenetic analyses of Nigerian S. intermedius with others from GenBank recover four lineages. Evidence of genetic structuring is consistent with geographic regions of sub-Saharan Africa. Thus, cryptic lineage diversity may illustrate species' adaptive responses to local environmental conditions. Finally, our study underscores the importance of incorporating morphology and DNA barcoding in species identification. Although developing a complete DNA barcode reference library for Nigerian ichthyofauna will facilitate species identification and diversity studies, taxonomic revisions of DNA sequences submitted in databases alongside voucher specimens are necessary for a reliable taxonomic and diversity inventory.

DNA barcoding discriminates freshwater fishes from southeastern Nigeria and provides river system-level phylogeographic resolution within some species.

BACKGROUND AND AIMS: Fishes are the main animal protein source for human beings and play a vital role in aquatic ecosystems and food webs. Fish identification can be challenging, especially in the tropics (due to high diversity), and this is particularly true for larval forms or fragmentary remains. DNA barcoding, which uses the 5' region of the mitochondrial cytochrome c oxidase subunit I (COI) as a target gene, is an efficient method for standardized species-level identification for biodiversity assessment and conservation, pending the establishment of reference sequence libraries. MATERIALS AND METHODS: In this study, fishes were collected from three rivers in southeastern Nigeria, identified morphologically, and imaged digitally. DNA was extracted, PCR-amplified, and the standard barcode region was bidirectionally sequenced for 363 individuals belonging to 70 species in 38 genera. All specimen provenance data and associated sequence information were recorded in the barcode of life data systems (BOLD; www.barcodinglife.org ). Analytical tools on BOLD were used to assess the performance of barcoding to identify species. RESULTS: Using neighbor-joining distance comparison, the average genetic distance was 60-fold higher between species than within species, as pairwise genetic distance estimates averaged 10.29% among congeners and only 0.17% among conspecifics. Despite low levels of divergence within species, we observed river system-specific haplotype partitioning within eight species (11.4% of all species). CONCLUSION: Our preliminary results suggest that DNA barcoding is very effective for species identification of Nigerian freshwater fishes.