Toxicity of 2-methyl-4-chlorophenoxy acetic acid alone and in combination with cyhalofop-butyl to Cyprinus carpio embryos.

Herbicides may pose considerable danger to non-target aquatic organisms and further threaten human health. The present investigation was aimed to assess the effects of 2-methyl-4-chlorophenoxy acetic acid (MCPA-Na) on Cyprinus carpio embryos. Embryos were exposed to six concentrations of MCPA-Na (0, 52, 54, 56, 58 and 60 mg/L) for 96 h. A series of symptoms were observed in developmental embryos during MCPA-Na exposure, including increased death, hatching inhibited and morphological deformities. Further, MCPA-Na exposure leading to a series of morphological changes (pericardial edema, tail deformation, and spine deformation) in embryos, which were consistent with modifications in the associated genes. In this work, we also investigated the joint toxicity of herbicides (MCPA-Na and cyhalofop-butyl) commonly used in paddy fields on carp embryos, using the 96 h-LC50 of herbicides (59.784 mg/L MCPA-Na and 1.472 mg/L cyhalofop-butyl) and confirmed that a synergistic effect existing in the binary mixtures.

Comparative transcriptome analyses and identification of candidate genes involved in vertebral abnormality of bighead carp Hypophthalmichthys nobilis.

Body deformity occurs both in wild and farmed fishes, which is one of the most challenging problems for aquaculture industry. In most cases, such body deformities are linked to skeletal deformities. Currently, very limited information is available on skeletal deformities of farmed fish species which may be caused by genetic factor. In this study, we performed muscle and vertebra transcriptome analyses in body deformity and normality of bighead carp Hypophthalmichthys nobilis (from one meiotic gynogenesis family) using RNA-Seq. A total of 43,923 and 44,416 unigenes were predicted in muscles and vertebrae, respectively. Based on these data, we further explored the gene expression profiles in gynogenetic normal and abnormal bighead carp. No differentially expressed gene (DEG) was found in transcriptome data of muscles. Totally, 20 key DEGs were identified in transcriptome data of vertebrae, such as low density lipoprotein-related protein 2 (lrp2), bone morphogenetic protein 2B (bmp2b) and collagen alpha-1(IV) (col4a1). 12 potential pathways were also identified in vertebra transcriptome data, which were mainly involved in development, growth, cytoskeleton and energy metabolism, such as MAPK signaling pathway, regulation of actin cytoskeleton and TGF-beta signaling pathway. Results of this study will be informative for the understanding of genetic mechanisms for body shape formation and also provide potential candidate genes for selection program involved in body shape and skeletal development in H. nobilis.

Ultrastructural Deformities in the Scales of Cyprinus carpio Inhabiting Two Government-Sponsored Fish Farms in North-East India Contaminated by Municipal Wastes and Other Pollutants.

Scanning electron microscopy of the scales of Cyprinus carpio, inhabiting two government-sponsored fish farms, with different sources of pollutants, in the North-East state of Meghalaya, India revealed extensive damage to them. Lepidonts, which are teeth-like structures present on the edge of circulii, were discovered to be either partly or completely damaged. The dislodgment of complete rows of lepidonts along with broken lepidonts was also observed. Loss of symmetry and torn areas of circulii are some of the abnormalities also observed as compared to control samples. The alterations observed could be because of the high sensitivity of the scales to environmental stress and their capacity to react to low concentrations of pollutants, which makes them a sensitive bioindicator of water pollution. The study hence not only represents an advance in pollution research, by employing a highly sophisticated analytical approach to reveal the above-mentioned microstructural scale abnormalities, but it can also have scientific impact on the society at large and the fish culturist in particular in understanding the destructive impact of pollution caused by mismanagement of city garbage and agricultural effluents.

The effects of Roundup on gametes and early development of common carp (Cyprinus carpio L).

To determine the effects of Roundup, a commercial formulation of glyphosate, gametes, and embryos of common carp (Cyprinus carpio L) was exposed to wide range of herbicide concentrations (0.0, 0.1, 0.5, 2.0, 5.0, 10.0, 20.0, and 50.0 mg/l). The obtained results showed different effects of Roundup on common carp gametes. Herbicide reduced swelling of eggs (but the effect was not concentration-related), while sperm showed low sensitivity to Roundup (time of spermatozoa motility was reduced in a significant way only at 20 mg/l, and at remaining concentrations, only a slight tendency was observed). During the embryonic development, Roundup caused a decrease of common carp embryonic survival (and the effect was concentration-related); however, it had no effect on development rate. During the embryogenesis, three types of embryo body malformation were observed: yolk sac edema, spine curvature, and shortening of body, but their frequencies were not associated with the presence or concentration of herbicide. However, Roundup affected quality of newly hatched larvae of common carp by increasing their mortality. No effect of herbicide on percentage of deformed larvae was observed but larvae hatched in water with Roundup tended to show more complex anomalies compared to those from the control. Obtained data showed that even low concentrations of this herbicide in waters can significantly reduce egg swelling, survival of embryos, and quality of fish larvae.

Case studies of spinal deformities in ornamental koi, Cyprinus carpio L.

This is a study of vertebral deformities in ornamental koi based on computed radiography and skeletons cleaned by dermestid beetles (Dermestes maculatus). All koi developed gradual onset of swimming abnormalities as adults. Extensive intervertebral osteophyte formation correlated with age of fish and was associated with hindquarter paresis in one koi. Vertebral compression and fusion were the most common spinal deformities occurring at multiple sites, similar to findings in other farmed fish. Site-specific spinal deformities were thought to develop due to differences in swimming behaviour and rates of vertebral growth. One koi had offspring with spinal deformities. Spinal deformities are significant problems in both European and Australian food fish hatcheries. The heritability of vertebral deformities in farmed fish is reportedly low unless there is concurrent poor husbandry or nutritional deficiencies. The specific aetiologies for vertebral deformities in koi in this study could not be ascertained. Current knowledge on spinal deformities in the better studied European food fish species suggests multifactorial aetiologies. Future research should include prospective longitudinal studies of larger numbers of koi from hatch and consideration of all potential risk factors such as husbandry, nutrition, temperature, photoperiod and genetics.

Avaliação da qualidade da água do Rio dos Sinos (Brasil) por meio do teste de micronúcleos em Cyprinus carpio e de análises físico-químicas e microbiológicas / Evaluation of water quality of the Sinos River (Brazil) by micronucleus test in Cyprinus carpio and physicochemical and microbiological analysis

O Rio dos Sinos está localizado na região sul do Brasil, possui aproximadamente 190 km de extensão e fornece água para atividades agrícolas, industriais e para o consumo de mais de um milhão de habitantes. Este rio é considerado um dos mais poluídos do Brasil e, portanto, estudos para avaliar a qualidade da água ao longo de sua extensão são necessários. O teste de micronúcleos em eritrócitos de peixes tem sido utilizado com sucesso para detectar a presença de poluentes mutagênicos nos ambientes aquáticos. O objetivo do presente estudo foi avaliar a qualidade da água do Rio dos Sinos por meio do teste de micronúcleos em Cyprinus carpio bem como de análises físico-químicas e microbiológicas. Foram coletadas amostras de água nos trechos superior (Caraá), médio (Parobé) e inferior (Novo Hamburgo) do Rio dos Sinos em dezembro de 2013. As amostras foram transportadas para o laboratório para análise de 14 parâmetros de qualidade da água bem como para a exposição de espécimes de C. carpio por 72h em aquários. Não foram verificadas diferenças significativas na frequência de micronúcleos e anormalidades nucleares nos grupos expostos a água do rio em comparação ao grupo controle. Apenas um parâmetro de qualidade da água foi observado em desacordo com a legislação no trecho superior do rio, enquanto que nos trechos médio e inferior foram seis e cinco parâmetros, respectivamente. Os resultados demonstram que o Rio dos Sinos apresenta redução da qualidade da água ao longo de sua extensão e indicam ausência de potencial genotóxico no período amostrado.

The Sinos River is located in South of Brazil, it has about 190 km of extension and provides water for agricultural and industrial activities, and for consumption of more than one million inhabitants. This river is considered one of the most polluted rivers in Brazil; therefore, studies aiming to evaluate the water quality along its course are necessary. The micronucleus test in fish erythrocytes has been successful used to detect mutagenic pollutants in the aquatic environment. The objective of this study was to evaluate the water quality of the Sinos River by the micronucleus test in Cyprinus carpio, as well as physicochemical and microbiological analyses. Water samples were collected in the upper (Caraá municipality), middle (Parobé municipality) and lower (Novo Hamburgo municipality) sections of the Sinos River, in December 2013. The samples were transported to the laboratory for analysis of 14 water quality parameters and for exposure of C. carpio for 72 hours in aquaria. Significant differences in micronucleus and nuclear abnormalities frequencies were not found between the control and the exposed groups. Only one parameter of water quality exceeded the limit of the legislation in the upper section of the river, while in the middle and lower sections, six and five parameters, respectively. These results show that the Sinos River presents a reduction in water quality along its extension and indicates the absence of genotoxic potential in the sampled period.

Phenolic endocrine-disrupting chemicals and intersex in wild crucian carp from Hun River, China.

This study investigated the concentrations of phenolic endocrine-disrupting chemicals (EDCs) and the incidence of ovo-testis (intersex) in wild crucian carp (Carassius carassius) sampled from the Hun River. As expected, nonylphenols (NP) and bisphenol A (BPA) were detected in all samples, and octylphenols (OP) were found in most samples. NP concentrations ranged from 1290 ± 584 to 3111 ± 2071 ng g(-1) wet weight (ww) in July and from 1132 ± 644 to 1556 ± 587 ng g(-1) ww in November; OP ranged from 6 ± 7 to 46 ± 38 ng g(-1) ww in July and from no detection to 22 ± 16 ng g(-1) ww in November; and BPA ranged from 4 ± 9 to 41 ± 24 ng g(-1) ww in July and from 6 ± 5 to 59 ± 24 ng g(-1) ww in November. Moreover, the concentrations of these compounds were higher in fish found down-stream of the sewage treatment plant (STP), and the lowest concentrations were found in fish up-stream of the STP. Concentrations of these EDCs in muscles might be correlated with the prevalence of intersex traits in wild fish, suggesting that these compounds contribute, at least in part, to the occurrence of intersex morphology. Thus, phenolic EDCs discharged from the STP pose a risk to the aquatic ecosystem in the Hun River.

Identification of some skeletal deformities in freshwater teleosts raised in Egyptian aquaculture.

This study examines different forms of skeletal deformities detected in fish species collected from two Egyptian aquaculture facilities during two harvest seasons in 2008. Various patterns of skeletal deformities were observed in 19 of 959 fish collected. Deformities were diagnosed using a number of techniques including clinical, radiographic, sonographic and histopathological evaluations. Observed deformities included: lordosis and kyphosis in African catfish (Clarius gariepenius); lateral projection of the mandible, parrot-like head, scoliosis, kyphosis, lordosis and fusion of dorsal with anal fins in Nile tilapia (Oreochromis niloticus); and stump body, scoliosis and mandibular joint deformity in common carp (Cyprinus carpio). Relative incidences of deformities in fish from a facility located in the Sharkia province were 5.12%, 2.66% and 2.85% among catfish, Nile tilapia and common carp, respectively. At a second fish farm located in the Kafr Elsheikh province, the incidences of deformities were 1.02%, 1.55% and 0% among catfish, Nile tilapia and common carp, respectively. Some of the deformities were confirmed using both sonographic and histopathological evaluations. The reasons for the observed deformities could not be definitively determined, but possible aetiologies are discussed.

Structural abnormalities of common carp Cyprinus carpio spermatozoa.

Spermatozoa of common carp Cyprinus carpio are typically consist of a primitive head without acrosome, a midpiece with several mitochondria, a centriolar complex (proximal and distal centriole), and one flagellum. During an evaluation of the motility of common carp spermatozoa, we found spermatozoa with more than one flagellum and/or "double head" in three different individuals. This may be related to abnormal spermatogenesis. Ultrastructure and physiological parameters of spermatozoa were examined using light microscopy (dark field with stroboscopic illumination), transmission and scanning electron microscopy, and flow cytometry. The recorded pictures and videos were evaluated using Olympus MicroImage software. All spermatozoa with more than one flagellum had a larger head and shorter flagella. They occasionally demonstrated several cytoplasmic channels separating the flagella from the midpiece. Each flagellum was based upon its own centriolar complex, with the connection of the flagellum to the head always at a constant angle. The flagella always consisted of nine peripheral pairs and one central doublet of microtubules. Sperm exhibited a relative DNA content similar to that found in sperm from normal males, with higher coefficients of variation. Although similar abnormalities have been found in livestock, where they were described as a defect in spermiogenesis, no comparable results have been reported in fish. The frequency at which these abnormalities occurs, the fertilization ability of males with defects in spermiogenesis, the influence of these abnormalities on progeny in terms of ploidy level, and the occurrence of deformities warrant further investigation.

The effects of heavy metals on embryonic development of fish (a review).

Early developmental stages of fish are particularly sensitive to water pollution. Heavy metals may affect various developmental processes during the embryonic period, which results in a reduction of offspring quantity and quality. Waterborne metals may accumulate in the gonads of spawners and adversely affect gamete production and viability, or exert direct toxic influence upon developing embryos. The egg shell does not fully protect the embryo against metal penetration, particularly during the swelling phase; thus, metals may accumulate in the egg. The results depend on metal concentration and range from developmental disturbances to death of the embryo. Metals disturb various processes of fish embryonic development and affect the development rate. Early stages just after fertilization are particularly sensitive to metal intoxication, when most disturbances and the highest embryonic mortality occur. Waterborne metals also promote developmental anomalies during organogenesis, including body malformations. Heavy metals often induce a delay in the hatching process, premature hatching, deformations and death of newly hatched larvae. All these disturbances result in reduced numbers and poor quality of the larvae, which show small body size, high frequency of malformations and reduced viability.

Gonadosomatic index and testis morphology of common carp (Cyprinus carpio) in rivers contaminated with estrogenic chemicals.

To study the effect of estrogenic chemicals on fish, the gonadosomatic index (GSI = [testis weight/body weight] x 100) and testis histology of mature common carp (Cyprinus carpio) from 2 contaminated sites (Ishizu and Wada rivers, Osaka) and a control site were examined between June 1998 and March 2001. The concentration of nonylphenol, bisphenol A and 17beta-estradiol in the Ishizu river was 3-4 times higher than in the Wada river. In the pre-breeding and breeding seasons, there were no significant differences in body weight among carp from the 3 sites, the body weight of Ishizu river carp being significantly lower (p<0.05) than that of Wada river fish only in the post-breeding season. The GSI and testis weight in fish from the Ishizu river were significantly lower (p<0.05) than in control fish during all phases of gonadal cycle and lower than in Wada river fish in the pre-breeding and post-breeding season. No histological abnormalities were found in the testes of the males examined. Histological observation of the testes revealed a delay in the onset of spermatogenesis in fish from the Ishizu river compared with those from the other sites. These results clearly imply that the estrogenic chemicals in the Ishizu river adversely affect the testis development of the fish.