The introduction of Physa acuta (Gastropoda: Physidae) on Ilha Grande, Southeast Brazil, from initial stages to an established population / A introdução de Physa acuta Draparnaud, 1805 (Gastropoda: Physidae) na Ilha Grande, Sudeste do Brasil, desde os estágios iniciais até uma população estabelecida

Abstract This study presents a four-year follow-up of an introduced population of Physa acuta Draparnaud, 1805, from initial stages to an established population. This introduction occurred on a small impacted stream of Vila do Abraão, the main village of Ilha Grande (Angra dos Reis, Rio de Janeiro, Brazil). The population size increased during the study, and presented a relationship to environmental factors, especially with rainfall. On the initial stages of introduction prevailed the smaller specimens, but on the overall, predominated the intermediate size classes. After less than a year, P. acuta becomes established on this stream and was possibly affecting the other species found on the stream. The information presented here is useful to understand the invasion process of invasive snails, as well as directing conservation efforts.

Resumo Neste estudo é apresentado um acompanhamento de quatro anos de uma população de Physa acuta Draparnaud, 1805, desde os estágios iniciais da introdução até seu pleno estabelecimento. Esta introdução ocorreu em um pequeno riacho impactado da Vila do Abraão, o principal vilarejo da Ilha Grande (Angra dos Reis, Rio de Janeiro, Brasil). A população cresceu durante o estudo, e apresentou uma relação com os fatores ambientais, especialmente a pluviosidade. Nos estágios iniciais de introdução prevaleceram os exemplares menores, contudo no panorama geral, predominaram as classes de tamanho intermediárias. Em menos de um ano de introdução, P. acuta se estabeleceu neste riacho, e possivelmente está afetando as demais espécies encontradas no rio. A informação apresentada aqui é importante para a compreensão do processo de invasão de moluscos invasores no Brasil, assim como no direcionamento de esforços de conservação.

Rab25 gene down-regulates snail expression and inhibits epithelial mesenchymal transformation of human breast cancer MDA-MB-231 cells / 中华内分泌外科杂志

Objective:To probe into Rab25 Gene’s Effect on TGF-β inhibition of proliferation, invasion and epithelial mesenchymal transformation (EMT) of breast cancer MDA-MB-231 cells and explore its molecular mechanism.Methods:The experiment was divided into three groups: control group,TGF-β Group and si-Rab25 group. TGF-β induced MDA-MB-231 cell model of EMT was built. CCK-8 assay was used to detect cell proliferation. Transwell assay was used to detect the ability of cell invasion and migration.Western blot was used to detect the changes of related proteins in each group.Results:After stimulating MDA-MB-231 cells with TGF-β, Rab25 gene was highly expressed. Compared with TGF-β group (57.48±%3.62%), the migration ability and invasion ability of cells in si-Rab25 group (33.49%±2.93%) decreased by 41.7%, with a significant difference ( P<0.05). Compared with TGF-β group (153.21%±4.17%), the proliferation ability of cells in si-Rab25 group (115.32%±5.69%) decreased by 24.73%, with a significant difference ( P<0.05). The expression of MDA-MB-231 fine EMT related protein in si-Rab25 group was significantly different from that in TGF-β group ( P<0.05). The expression of p-AKT and Snail protein in si-Rab25 group was significantly lower than that in TGF-β group ( P<0.05) . Conclusions:Rab25 gene is highly expressed in MDA-MB-231 cells. Silencing Rab25 gene can activate AKT signal pathway, inhibit Snail protein expression, regulate EMT related protein expression, and inhibit EMT transformation.

Berberine Promotes Mouse Breast Cancer Metastasis to Lung via Inducing Epithelial–mesenchymal Transition / 肿瘤防治研究

Objective To study the effect and mechanism of berberine (BBR) on the lung metastasis of mouse breast cancer via epithelial-mesenchymal transition (EMT). Methods CCK-8 and Transwell migration assays were utilized to investigate the proliferation and migration properties of breast cancer 4T1 cells after BBR treatment.Mouse 4T1-Luc cells were injected into mice under the fourth mammary fat pad, and the mice were then randomly divided into the control and BBR groups.The mice in the BBR group received daily intraperitoneal injections of BBR working solution and those in the control group were continuously intraperitoneally injected with the same volume of the solvent used to dissolve BBR powder.Tumor metastasis in the lungs of living mice was detected by using an in vivo imaging system.After 42 days of administration, lung metastasis was measured via microscopy and HE staining.Western blot analysis was used to examine the effects of BBR on the expression of EMT-related proteins (Vimentin and Snail) as well as the activation of the Akt and ERK signaling pathways. Results BBR significantly promoted 4T1 cell migration (P < 0.05).In vivo experiments showed that the number of lung metastases in the BBR group had significantly increased compared with that in control group (P < 0.05) as observed under microcopy and histological staining.Compared with the control group, BBR upregulated the expression levels of Vimentin and Snail as well as the phosphorylated levels of p-Akt and p-ERK (P < 0.05). Conclusion BBR may promote EMT and lung metastasis of breast cancer 4T1 cells by activating the expression of proteins in the p-Akt and p-ERK pathways.

Considering the role of the assassin snail Anentome helena as a biological control of Bithynia siamensis goniomphalos, the first intermediate host of Opisthorchis viverrini

This study aims to investigate whether the assassin snail Anentome helena may serve as a biological control agent of Bithynia siamensis goniomphalos, the 1st intermediate host of Opisthorchis viverrini. Experiments were carried out in the laboratory, and the results found that A. helena showed the variation and selection of mollusc prey. A. helena can consume B. siamensis goniomphalos, which is remarkable because this snail can compete with other snails and could be used as a biological control. The consumption rate of the predator was compared, and it was found that A. helena prefers to consume Indoplanorbis exustus, followed by Pomacea canaliculata, Melanoides tuberculata, Filopaludina sumatrensis speciosa, Lymnaea sp., and B. siamensis goniomphalos. This is the first report of an experimental study controlling B. siamensis goniomphalos using A. helena. Our data imply that A. helena can control the B. siamensis goniomphalos population with good results, especially in the absence of other snail species.

The introduction of Physa acuta (Gastropoda: Physidae) on Ilha Grande, Southeast Brazil, from initial stages to an established population / A introdução de Physa acuta Draparnaud, 1805 (Gastropoda: Physidae) na Ilha Grande, Sudeste do Brasil, desde os estágios iniciais até uma população estabelecida

This study presents a four-year follow-up of an introduced population of Physa acuta Draparnaud, 1805, from initial stages to an established population. This introduction occurred on a small impacted stream of Vila do Abraão, the main village of Ilha Grande (Angra dos Reis, Rio de Janeiro, Brazil). The population size increased during the study, and presented a relationship to environmental factors, especially with rainfall. On the initial stages of introduction prevailed the smaller specimens, but on the overall, predominated the intermediate size classes. After less than a year, P. acuta becomes established on this stream and was possibly affecting the other species found on the stream. The information presented here is useful to understand the invasion process of invasive snails, as well as directing conservation efforts.

Neste estudo é apresentado um acompanhamento de quatro anos de uma população de Physa acuta Draparnaud, 1805, desde os estágios iniciais da introdução até seu pleno estabelecimento. Esta introdução ocorreu em um pequeno riacho impactado da Vila do Abraão, o principal vilarejo da Ilha Grande (Angra dos Reis, Rio de Janeiro, Brasil). A população cresceu durante o estudo, e apresentou uma relação com os fatores ambientais, especialmente a pluviosidade. Nos estágios iniciais de introdução prevaleceram os exemplares menores, contudo no panorama geral, predominaram as classes de tamanho intermediárias. Em menos de um ano de introdução, P. acuta se estabeleceu neste riacho, e possivelmente está afetando as demais espécies encontradas no rio. A informação apresentada aqui é importante para a compreensão do processo de invasão de moluscos invasores no Brasil, assim como no direcionamento de esforços de conservação.

The introduction of Physa acuta (Gastropoda: Physidae) on Ilha Grande, Southeast Brazil, from initial stages to an established population

Abstract This study presents a four-year follow-up of an introduced population of Physa acuta Draparnaud, 1805, from initial stages to an established population. This introduction occurred on a small impacted stream of Vila do Abraão, the main village of Ilha Grande (Angra dos Reis, Rio de Janeiro, Brazil). The population size increased during the study, and presented a relationship to environmental factors, especially with rainfall. On the initial stages of introduction prevailed the smaller specimens, but on the overall, predominated the intermediate size classes. After less than a year, P. acuta becomes established on this stream and was possibly affecting the other species found on the stream. The information presented here is useful to understand the invasion process of invasive snails, as well as directing conservation efforts.

Resumo Neste estudo é apresentado um acompanhamento de quatro anos de uma população de Physa acuta Draparnaud, 1805, desde os estágios iniciais da introdução até seu pleno estabelecimento. Esta introdução ocorreu em um pequeno riacho impactado da Vila do Abraão, o principal vilarejo da Ilha Grande (Angra dos Reis, Rio de Janeiro, Brasil). A população cresceu durante o estudo, e apresentou uma relação com os fatores ambientais, especialmente a pluviosidade. Nos estágios iniciais de introdução prevaleceram os exemplares menores, contudo no panorama geral, predominaram as classes de tamanho intermediárias. Em menos de um ano de introdução, P. acuta se estabeleceu neste riacho, e possivelmente está afetando as demais espécies encontradas no rio. A informação apresentada aqui é importante para a compreensão do processo de invasão de moluscos invasores no Brasil, assim como no direcionamento de esforços de conservação.

Optimization of the medium and fermentation condition for the Penicillium aurantiocandidum Z12 strain with molluscicidal actions against Oncomelania hupensis / 中国血吸虫病防治杂志

Objective To optimize the culture and fermentation conditions of the Penicillium aurantiocandidum Z12 strain, a fungal strain with molluscicidal actions against Oncomelania hupensis, so as to provide the basis for the research and development of molluscicidal active substances from the P. aurantiocandidum Z12 strain and its fermentation broth and large-scale fermentation. Methods The carbon source, nitrogen source and mineral salts were identified in the optimal culture medium for the P. aurantiocandidum Z12 strain with a single-factor experiment to determine the best fermentation condition for the P. aurantiocandidum Z12 strain. Factors that significantly affected the growth of the P. aurantiocandidum Z12 strain were identified using the Plackett-Burman design, and the best range of each factor was determined using the steepest climb test. Response surface analyses of temperature, pH value, seeding amount and liquid-filling quantity were performed using the Box-Behnken design to create a regression model for fermentation of the P. aurantiocandidum Z12 strain to identify the optimal culture medium. Results Single-factor experiment preliminarily identified the best culture medium and conditions for the P. aurantiocandidum Z12 strain as follows: sucrose as the carbon source at approximately 20 g/L, tryptone as the nitrogen source at approximately 5 g/L, K2HPO4 as the mineral salt at approximately 5 g/L, initial pH at approximately 8, temperature at approximately 28 °C, seeding amount at approximately 6%, and liquid-filling quantity at approximately 50 mL/100 mL. Plackett-Burman design showed that factors that significantly affected the growth of the P. aurantiocandidum Z12 strain included temperature (t = −5.28, P < 0.05), seeding amount (t = 5.22, P < 0.05), pH (t = −4.30, P < 0.05) and liquid-filling quantity (t = −4.39, P < 0.05). Steepest climb test showed the highest mycelial growth at pH of 7.5, seeding amount of 8%, and liquid-filling quantity of 40 mL/100 mL, and this condition was selected as the central point of response surface analysis for the subsequent optimization of fermentation conditions. Response surface analyses using the Box-Behnken design showed that the optimal conditions for fermentation of the P. aurantiocandidum Z12 strain included sucrose at 15 g/L, tryptone at 5 g/L, K2HPO4 at 5 g/L, temperature at 28.2 °C, pH at 7.5, seeding amount at 10%, and liquid-filling quantity at 35.8 mL/100.0 mL, resulting in 0.132 g yield of the P. aurantiocandidum Z12 strain. Conclusion The optimal culture condition for the P. aurantiocandidum Z12 strain has been identified, and the optimized culture medium and fermentation condition may effectively improve the fermentation yield of the P. aurantiocandidum Z12 strain.

Monitoring and analysis of endemic situation of schistosomiasis in Suzhou New District from 2004 to 2021 / 上海预防医学

ObjectiveTo analyze the monitoring data of schistosomiasis from 2004 to 2021 in Suzhou New District， Jiangsu Province， and to provide evidence for improving schistosomiasis elimination strategies. MethodsFollowing the Opinions on Prevention and Control of Schistosomiasis， Parasitic Diseases and Endemic Diseases in Suzhou and the Technical Plan for Prevention and Control of Schistosomiasis， Parasitic Diseases and Endemic Diseases in Suzhou， the monitoring of schistosomiasis in the population and snail habitats from 2004 to 2021 was conducted. The Mann-Kendall method and Joinpoint regression method were employed to analyze the trend of epidemic indicators （such as seropositive rate， prevalence of snail frames， etc.）. Time series analysis （exponential smoothing model） was conducted to predict snail occurrence. ResultsFrom 2004 to 2021， a total of 73 680 people were serologically tested for schistosomiasis， with a positive rate of 0.084%. The seropositivity rate showed statistically significant differences between different years （χ2=70.73， P<0.05）， but there was no significant trend over time. In addition， 3 053 fecal tests were conducted and no positive result was found. The snail habitats covered an area of 70.11 hm2 and showed a decreasing trend （Z=-1.97， P<0.05）. A total of 30 093 frames were surveyed， of which 19.038% contained snails. The difference in the prevalence of snail frames between different years was statistically significant （χ2=7 203.09， P<0.05）， with a decreasing trend in the prevalence of snail frames （Z=-2.05， P<0.05）. A total of 26 296 live snails were seized and density of live snails was 0.874 snails per frame， showing a decreasing trend in the density of live snails （Z=-2.35， P<0.05）. A total of 12 391 snails were dissected and no infected snail was found. The areas treated with molluscicides remained stable at 264.60 hm2. An area of 27.77 hm2 achieved the goal of snail eradication through environmental modification， with a decreasing trend （Z=-2.44， P<0.05）. It is estimated that the prevalence of snail frames and snail density will remain relatively stable from 2022 to 2026， but the snail habitat area will fluctuate significantly， showing an increasing trend. ConclusionNo indigenous cases of schistosomiasis and no infected snails are reported， indicating the successful consolidation of schistosomiasis prevention and control measures. However， the snail habitat area fluctuates greatly with an increasing trend， suggesting the need for long-term Oncomelania snail monitoring in local areas.

Estudo da imunoexpressão de proteínas envolvidas na transição epitélio-mesênquima em tumores de glândula salivar / Study of the immunoexpression of proteins involved in the epithelial-mesenchymal transition in salivary gland tumors

Os tumores de glândula salivar (TGS) apresentam notável complexidade clínica e biológica, razão para a qual muitos estudos investigam os eventos envolvidos na sua progressão. Uma das dinâmicas envolvidas na invasão tumoral de diversos tipos de carcinomas é a transição epitélio-mesênquima (TEM). Neste processo, as células epiteliais sofrem transição para um estado mesenquimal móvel, favorecendo a invasão e metástase. Sendo assim, esta pesquisa analisou a expressão imuno-histoquímica de E-caderina, Twist1, Snail1, α-SMA, metaloproteinases de matriz 9 (MMP-9) e Vimentina (VM) em 90 casos de TGS, correlacionando-os entre si e com parâmetros clinicopatológicos. Foram selecionados 20 casos de Adenoma pleomórfico (AP), 20 casos de Carcinoma mucoepidermoide (CME), 20 casos de Carcinoma adenoide cístico (CAC), 10 casos de Adenocarcinoma polimorfo (ACP), 10 casos de Carcinoma epitelial-mioepitelial (CEME) e 10 casos de Carcinoma ex-adenoma pleomórfico (CexAP). A análise de E-caderina, Twist1, Snail1 foi realizada em parênquima tumoral sendo observado o percentual de células positivas (PP), com escores variando de 0 a 4, e a intensidade de expressão (IE), cujos escores variaram de 0 a 3. A avaliação de MMP-9 foi realizada em parênquima e estroma tumoral, também avaliando-se a PP e a IE, ambos baseados em escores que variaram de 0 a 3. A marcação para α-SMA e VM foi analisada em região de estroma tumoral. Células positivas para α-SMA foram contabilizadas em 10 campos, obtendo-se, então a média. A VM foi avaliada de forma qualitativa, utilizando-se 4 escores de acordo com a IE e se a marcação é difusa ou focal. Os dados obtidos foram analisados no software Statistical Package for Social Science, GraphPad Prism e STATA. O nível de significância de 5% foi adotado para os testes estatísticos. Foi verificada menor imunomarcação de E-caderina nos APs em relação às neoplasias malignas de glândula salivar (NMGS). Observou-se baixa imunoexpressão de Twist1 e Snail1 em APs. Em relação a expressão nuclear do Twist1, constatou-se maior expressão nas neoplasias malignas quando comparadas aos APs. Ainda, Twist1 em núcleo foi correlacionado à expressão citoplasmática de E-caderina nas NMGS. No que concerne aos parâmetros clinicopatológicos, esta proteína se relacionou estatisticamente com maiores chances de óbito. Foi evidenciada baixa imunoexpressão de Snail1 entre as NMGS. No entanto, na análise dos CACs, foi verificada maior expressão nuclear na variante sólida em relação às demais. A expressão de MMP-9 em parênquima demonstrou correlação positiva com Twist1 citoplasmático e Snail1nuclear nas NMGS. A MMP-9 também apresentou correlação positiva na comparação da sua imunoexpressão em região de parênquima e de estroma. A VM se apresentou como um biomarcador a ser considerado na avaliação clínica dos pacientes, já que esta apresentou relação significativa com tamanho do tumor (T3-T4) e maior frequência de óbito. Ademais, a alta expressão desta proteína se apresentou como um fator preditivo independente para piores taxas de sobrevida global (SG). A avaliação dos demais fatores clinicopatológicos apresentou estágios clínicos avançados como indicador de valor prognóstico independente para menores taxas de SG, enquanto que para a sobrevida livre da doença, estes foram a localização em glândula salivar menor e presença de metástase à distância. Os resultados deste estudo sugerem que o processo de TEM pode estar relacionado ao estágio de diferenciação celular em APs e à progressão tumoral nas NMGS. Ressalta-se, também, maior participação de Twist1 e MMP-9 no cenário da TEM em tumores malignos de glândula salivar, além da possibilidade de utilização da VM como indicador de valor prognóstico (AU).

Salivary gland tumors (SGTs) present remarkable clinical and biological complexity; therefore, many studies investigate the events involved in their progression. One of the dynamics involved in the tumor invasion of different types of carcinomas is the epithelial-mesenchymal transition (EMT). In this process, epithelial cells undergo a transition to a mobile mesenchymal state, favoring invasion and metastasis. Therefore, this research analyzed the immunohistochemical expression of E-cadherin, Twist1, Snail1, α-SMA, vimentin (VM) and matrix metalloproteinase 9 (MMP-9) in 90 SGTs cases; correlations among the biomarkers, as well as between the biomarkers and clinicopathological parameters were made. We selected 20 cases of pleomorphic adenoma (PA), 20 cases of mucoepidermoid carcinoma (MEC), 20 cases of adenoid cystic carcinoma (ACC), 10 cases of polymorphous adenocarcinoma (PAC), 10 cases of epithelial-myoepithelial carcinoma (EMC) and 10 cases of carcinoma ex-pleomorphic adenoma (CXPA). E-cadherin, Twist1, and Snail1 were analyzed in tumor parenchyma, observing the percentage of positive cells (PP) using scores ranging from 0 to 4, and the expression intensity (EI), whose scores were ranged from 0 to 3. The evaluation of MMP-9 was performed in tumor parenchyma and stroma, also evaluating PP and IE, both based on scores that ranged from 0 to 3. The labeling for α-SMA and VM was analyzed in stromal cells. Positive cells for α-SMA were counted in 10 fields and the mean was calculated. VM was evaluated qualitatively, using 4 scores according to EI and whether the labeling was diffuse or focal. Obtained data were analyzed using Statistical Package for Social Science, GraphPad Prism, and STATA software. The significance level of 5% was adopted for the statistical tests. Patients were mostly female, with a mean age of 49.8 years; the major salivary glands were the most affected anatomical site, mainly the parotid gland. A lower E-cadherin immunostaining was verified in PAs in comparison to malignant neoplasms of salivary glands (MNSGs). Low immunoexpression of Twist1 and Snail1 was observed in PAs. Regarding the nuclear expression of Twist1, it was found greater expression in malignant neoplasms than in PAs. Furthermore, Twist1 in the nucleus was correlated with cytoplasmic expression of E-cadherin in MNSGs. Regarding clinicopathological parameters, this protein was statistically related to higher chances of death. Low immunoexpression of Snail1 was evidenced among the MNSGs. However, in the analysis of CACs, greater nuclear expression was observed in the solid variant compared to the others. Expression of MMP-9 in parenchyma showed a positive correlation with cytoplasmic Twist1 and Snail1nuclear in MNSGs. MMP-9 also showed a positive correlation when comparing its immunoexpression in the parenchyma and the stroma. VM was presented as a biomarker to be considered in the clinical evaluation of patients since it showed a significant correlation between greater tumor size and a higher frequency of death. Furthermore, the high expression of this protein appeared as an independent predictive factor for worse overall survival (OS) rates. The evaluation of the rest of the clinicopathological factors showed advanced clinical stages as an indicator of independent prognostic value for lower rates of OS. For disease-free survival, these indicators were the location in the minor salivary gland and the presence of distant metastasis. Our results suggest that the EMT may be related to myoepithelial differentiation in PAs and tumor progression in MNSGs. Also, Twist1 and MMP-9 appear to play a greater role in the scenario of EMT in MNSGs; finally, VM might be used as a prognostic value indicator (AU).

Low-dose cisplatin exposure and SNAIL, Vimentin, E-cadherin expression in HEPG2 cell line / Exposición a dosis bajas de cisplatino y expresión de SNAIL, Vimentina, E-cadherina en línea celular HEPG2

Cisplatin, the first platinum compound approved for cancer treatment, is widely used in the treatment of various cancers including hepatocellular carcinoma (HCC). HCC incidence rates rise globally. Epithelial mesenchymal transition (EMT) is implicated in cancer invasion and metastasis, which are associated with increased mortality. Cisplatin dose might influence cancer invasion and metastatic behavior of the cells. The aim of the study was to investigate the effect of low-dose cisplatin treatment on EMT- related changes in HepG2 cells. Following treatment with 4 µM cisplatin, HepG2 cells were evaluated morphologically. Gene expression of E-cadherin, Vimentin, Snail1 was assessed by quantitative PCR. Immunofluorescence analyses of NA-K ATPase were performed. Although the low-dose cisplatin treated cells exhibited a more stretched morphology, no statistical difference was detected in gene expression of E-cadherin, Vimentin, Snail1 and immunofluorescence of NA-K ATPase. Findings on low-dose cisplatin effects in HepG2 might contribute to the knowledge of antineoplastic inefficacy by further understanding the molecular mechanisms of drug action.

El cisplatino, el primer compuesto de platino aprobado para el tratamiento del cáncer, es ampliamente utilizado en el tratamiento de varios tipos de cáncer, incluido el carcinoma hepatocelular (CHC). Las tasas de incidencia de CHC aumentan a nivel mundial. La transición mesenquimal epitelial (EMT) está implicada en la invasión del cáncer y la metástasis, que se asocian con un aumento de la mortalidad. La dosis de cisplatino podría influir en la invasión del cáncer y el comportamiento metastásico de las células. El objetivo del estudio fue investigar el efecto del tratamiento con dosis bajas de cisplatino en los cambios relacionados con la EMT en las células HepG2. Tras el tratamiento con cisplatino de 4 µM, se evaluaron morfológicamente las células HepG2. La expresión génica de E-cadherina, vimentina, caracol1 se evaluó mediante PCR cuantitativa. Se realizaron análisis de inmunofluorescencia de NA-K ATPasa . Aunque las células tratadas con cisplatino en dosis bajas exhibieron una morfología más estirada, no se detectaron diferencias estadísticas en la expresión génica de E-cadherina, vimentina, Snail1 e inmunofluorescencia de NA-K ATPasa. Los hallazgos sobre los efectos del cisplatino en dosis bajas en HepG2 podrían contribuir al conocimiento de la ineficacia antineoplásica al comprender mejor los mecanismos moleculares de la acción del fármaco.

Waste leachate induces histological alterations in Limicolaria aurora (Jay, 1839) digestive gland

Unsanitary open dumping and burning of wastes in dumpsites results to constant leaching of waste pollutants into the surrounding environment with attendant harmful consequences on wildlife and public health. Here, we investigated the histological effects of Onitsha municipal waste leachate on the Giant African land snail, Limicolaria aurora (Jay 1839). The histopathological effects of waste leachate on the digestive cells of the snails were examined following standard protocols. Snails were exposed to different concentrations (0, 6.25, 12.5, 25.0 and 50.0%) of the leachate for a total duration of 21 days. Histological results obtained showed that waste leachate caused dose and duration dependent alterations in the digestive glands of L. aurora. Obvious histological variations were observed in snails exposed to 50% concentrations of the leachate compared to other doses. Observed histological changes were mainly degeneration of the digestive tubules, fragmentation and disappearance of the digestive cells, marked increase in excretory cells, necrosis and epithelia sloughing. This study therefore highlights the potential toxicity of waste leachate of causing mild to severe damage in the tissues of organisms. Reinforcing the fact, that waste leachate contains a mix of toxic substances that could pose a severe injury to the biological system of surrounding biota, including humans.

Prevalencia de Fasciola hepatica y Paramphistomidae en bovinos de doble propósito en una hacienda del trópico bajo andino colombiano / Prevalence of Fasciola hepatica and Paramphistomidae in dual purpose cattle in a farm in the Colombian low Andean tropic

RESUMEN En muchos países la fasciolosis y la paramfistomosis representan un grave problema para la salud del sector pecuario. En Colombia hay registros de ambas distomatosis en bovinos, la mayoría en el trópico alto andino, las cuales generan pérdidas econômicas anuales cercanas a 40 000 000 000 COP. El objetivo de esta investigación fue determinar la prevalencia de fasciolosis y paramfistomosis en vacunos de la hacienda La Candelaria, Caucasia (Colombia), y la presencia de caracoles hospederos intermediarios. Para cumplirlo, se realizó un estudio descriptivo de corte transversal con muestreo no probabilístico por conveniencia. Se recolectaron heces de los animales y se hizo el diagnóstico mediante la técnica modificada de Dennis. Se establecieron las prevalencias de los digeneos de acuerdo con el sexo, peso, edad y raza. Se recolectaron caracoles dulciacuícolas en la zona estudiada y se identificaron por morfologia. Se analizaron 466 muestras fecales de 178 bovinos, de las razas Cebú (Bos indicus), BON (blanco orejinegro) y del cruce entre ellas. Se diagnosticaron F. hepatica y Paramphistomidae con prevalencias del 2,2% y 30,9%, respectivamente. En el 1,1% de los vacunos se diagnosticó coinfección. Ambas trematodosis prevalecieron en las hembras (p = 0,03). Se identificaron moluscos dulciacuícolas Ampullariidae, Physidae y Planorbidae sin estadios larvarios de digeneos. Se concluyó que los bovinos de doble propósito de la hacienda La Candelaria están expuestos a F. hepatica y Paramphistomidae, y, probablemente, se infectan en los predios de la hacienda. Paramphistomidae es más prevalente que F. hepatica, lo cual concuerda con lo descrito en algunos estudios realizados en hatos del trópico alto andino colombiano. Paramphistomidae se encontró en todos los grupos etarios.

ABSTRACT Fasciolosis and paramphistomosis are a major health problem for the livestock economy worldwide. In Colombia, both distomatosis are reported in cattle, particularly in high Andean tropics, with annual economic losses close to COP 40 billion. The goal of this study was to determine the prevalence of fasciolosis and paramphistomosis in cattle from La Candelaria farm, Caucasia (Colombia), and the presence of intermediate host snails. A descriptive cross-sectional study was carried out with non-probability convenience sampling. Stool samples were collected from the animals and the diagnosis was made using the modified Dennis technique. Digenea prevalence were determined according to sex, weight, age, and race. Freshwater snails were collected in the studied area and were identified by morphology. 400 and 66 fecal samples from 178 bovines of the Zebu (Bos indicus), BON (white-eared white) breeds and the cross between them were analyzed. Fasciola hepatica and Paramphistomidae were diagnosed with a prevalence of 2,2% and 30,9%, respectively. Coinfection was diagnosed in 1,1% of the cattle. Both trematodosis were most frequent in females (p = 0,03). Freshwater molluscs Ampullariidae, Physidae and Planorbidae without digenea larval stages were identified. It was concluded that dual-purpose cattle from La Candelaria farm are exposed to F. hepatica and Paramphistomidae and are probably infected on the farm grounds. Paramphistomidae was more prevalent than F. hepatica, which agrees with other studies in herds from the Colombian high Andean tropics. Paramphistomidae was found in all age groups.

Relative condition factor and predictive model for the presence of the invasive snail Achatina (Lissachatina) fulica in Sergipe, Northeast Brazil / Fator de condição relativo e modelo preditivo para a presença do caracol invasor Achatina (Lissachatina) fulica em Sergipe, Nordeste do Brasil

Abstract: Achatina fulica is among the world's 100 most impactive invasive species, and is now found in almost all Brazilian states, including Sergipe. This exotic snail is known to have negative impacts, not only on the environment, due primarily to the rapid growth of its populations, but also on public health, given that it is an intermediate host of nematodes that cause zoonotic diseases. However, relatively little is known of the development of this snail, including its relative condition factor. We investigated the occurrence of A. fulica in 24 municipalities distributed in the eight subregions of the state of Sergipe in the dry and rainy season. Furthermore, we present here a predictive model for the occurrence of A. fulica based on the variation in climate and soil chemistry. This snail was more frequent on soils with a pH of 6.5-7.5. A negative correlation was found between the growth of A. fulica and the soil pH, then, the more acidic the soil, the more allometric the growth of A. fulica. The relative condition factor indicated differences in the development pattern of A. fulica among the eight subregions. The influence of rain in increasing the frequency of A. fulica showed a significant correlation. As well, higher temperatures influenced the resting behavior of A. fulica. The mathematical model used to identify the potential presence of A. fulica presented a high degree of agreement. This is the first ecological study of A. fulica to verify the association between the body mass-length relationship and the relative condition factor, and the results indicate that the development of this exotic land snail in Sergipe is influenced by climatic factors and principally, the soil pH. The predictive mathematical model provides valuable insights into the biotic and abiotic factors associated with the presence of A. fulica, and the influence of climatic variables and the chemical parameters of the soil on the occurrence of this species. These findings provide important guidelines for the development of measures for the control of A. fulica populations, which will contribute to both public and environment health.

Resumo: Achatina fulica está entre as 100 das piores espécies invasoras em todo o mundo, e no Brasil está presente em quase todos os estados, incluindo Sergipe. Este caracol exótico é conhecido por ter impactos negativos, não só no meio ambiente, devido principalmente ao rápido crescimento de suas populações, mas também na saúde pública, uma vez que é um hospedeiro intermediário de nematodeos causadores de doenças zoonóticas. No entanto, pouco se sabe a respeito do desenvolvimento dessa espécie, incluindo o fator de condição relativo. Investigamos a ocorrência de A. fulica em 24 municípios distribuídos nos oito territórios do estado de Sergipe no período seco e chuvoso. Além disso, apresentamos aqui um modelo preditivo para a ocorrência de A. fulica baseado na variação do clima e da química do solo. Esse caracol é mais frequente no solo com pH de 6,5 à 7,5, sendo esse padrão ideal para o crescimento isométrico. O crescimento alométrico de A. fulica apresentou correlação negativa com o pH do solo, quanto mais ácido for o solo, maior será o crescimento de A. fulica. O fator de condição de A. fulica, apresentou diferença no desenvolvimento nos oito Territórios. A influência da chuva na frequência de A. fulica apresentou correlação significativa. Além disso, temperaturas mais elevadas influenciaram no comportamento de repouso de A. fulica. O modelo matemático para identificar a possível presença de A. fulica apresentou uma concordância forte. Este é o primeiro estudo ecológico de A. fulica a verificar a associação entre a relação massa-comprimento e o fator de condição relativo, sendo possível evidenciar que essa espécie exótica em Sergipe sofre alterações no desenvolvimento, por fatores climáticos e principalmente pelo pH do solo. O modelo matemático preditivo fornece informações valiosas sobre os fatores bióticos e abióticos associados à presença de A. fulica e a influência de variáveis climáticas e dos parâmetros químicos do solo na ocorrência desta espécie. Esses achados fornecem importantes diretrizes para o desenvolvimento de medidas de controle de populações de A. fulica, que poderão contribuir para a saúde pública e ambiental.

Damping the Impact of Intraspecific Variability of Whorl Growth-rate on the Range of Intraspecific Variation of Adult Shell-size in Gastropods with Determinate Growth: Evidence of a Finely-tuned Regulating Process

Many species among shelled gastropods � either land snails or marine snails � exhibit determinate growth and, therefore, are expected to implement some regulating process aiming at limiting the impact on the final (adult) shell-size of the (possibly excessive) intraspecific variability in the rate of shell-development. Indeed, a usually more or less limited range of variation is allowed for adult shell-size, in those species having determinate growth. Mollusks are expected to be no exception in this respect and, up to now, the occurrence of such a regulating process has actually been reported systematically, despite a still too limited number of investigations. Yet, the question remained of the ability of this regulatory process to finely adjust the end of the final development of both body and shell (specifically in term of the final number of whorls), so as to limit the variations in adult shell-size, despite the amplitude of intraspecific variability in shell development rate. I provide here preliminary empirical evidence for such a 揻inely tuned� regulating process � the strength of which is mirrored by the degree of 揋ouldian� negative covariance between whorl growth-rate and the number of whorls at adulthood. As expected, the strength of the regulating process reveals systematically increasing (i) with the amplitude of the intraspecific variability in shell development-rate and (ii) with the narrowness of the range of variation allowed for the adult shell-size (narrowness which depends upon the identity of the particular species under consideration). In addition to the already reported usual occurrence of this regulating process among shelled gastropods, its 揻inely tuned� character provides here still further evidence as regards its major importance in controlling the species-specific adult body mass, and this even for typically soft-bodied animals.

Isolation and characterization of five novel disulfide-poor conopeptides from Conus marmoreus venom

Background: Conopeptides from cone snail venom have aroused great interest related to the discovery of novel bioactive candidates, due to their excellent prospects for the treatment of various health problems such as pain, addiction, psychosis and epilepsy. In order to explore novel biopeptides, we investigated the structure and function of five novel conopeptides isolated from the venom of Conus marmoreus from South China Sea. Methods: C. marmoreus crude venom was prepared, fractionated and purified by HPLC system. The primary sequences of the five novel disulfide-poor conopeptides Mr-1 to Mr-5 were identified by comprehensive analysis of de novo MALDI-TOF tandem mass spectrometry and Edman degradation data. In order to investigate their function, these five conopeptides were synthesized by Fmoc-SPPS chemistry, and their biological effects at several heterologous rat nicotinic acetylcholine receptor (nAChR) subtypes (α1β1δε, α3β2, α3β4, α4β2) were determined by electrophysiological technique. Results: Five novel disulfide-poor conopeptides were identified and named as follows: Mr-1 (DWEYHAHPKPNSFWT), Mr-2 (YPTRAYPSNKFG), Mr-3 (NVIQAPAQSVAPP NTST), Mr-4 [KENVLNKLKSK(L/I)] and Mr-5 [NAVAAAN(L/I)PG(L/I)V]. None of them contains a disulfide bond. The sequences of conopeptides Mr-2 to Mr-5 do not belong to any category of the known disulfide-poor conopeptides. No significant activity against the above nAChR subtypes were observed for the five conopeptides at 100 µM. Conclusion: We purified and structurally characterized five novel disulfide-poor conopeptides from C. marmoreus crude venom and first investigated their nAChR inhibitory effects. This work expanded our knowledge on the structure and function of disulfide-poor conopeptides from this cone snail venom.(AU)

First report of invasive Pomacea snails in Shandong Province / 中国血吸虫病防治杂志

Objective To characterize the species of invasive Pomacea snails that were discovered for the first time in Shandong Province. Methods Pomacea snails samples were collected in the field of Jining City, Shandong Province on October 2021 for morphological identification. Pomacea snails were randomly sampled and genomic DNA was extracted from foot muscle tissues of Pomacea snails for multiplex PCR amplification. The PCR amplification product was sequenced. Then, the sequence was aligned and a phylogenetic tree was created using the software MegAlign 7.1.0. In addition, Angiostongylus cantonensis infection was detected in Pomacea snails with the lung microscopy. Results A total of 104 living Pomacea snails were collected, and all were characterized as Pomacea spp. based on morphological features. Of 12 randomly selected adult Pomacea snails, multiplex PCR assay and sequencing identified eleven snails as P. canaliculata and one as P. maculata. No A. cantonensis infection was detected in 104 Pomacea snails. Conclusion This is the first report of invasive Pomacea snails in Shandong Province, where P. canaliculata and P. maculata are found.

Molluscicidal effect of 25% wettable powder of pyriclobenzuron sulphate in hilly schistosomiasis-endemic regions / 中国血吸虫病防治杂志

Objective To evaluate the molluscicidal effect of 25% wettable powder of pyriclobenzuron sulphate (WPPS) against Oncomelania snails in hilly schistosomiasis-endemic regions and test its toxicity to fish. Methods In October 2020, a snail-infested setting which had been cleared was selected in Nanjian County, Yunnan Province and divided into several blocks, and the natural snail mortality was estimated. 25% WPPS was prepared into solutions at concentrations of 1 and 2 g/L, and 25% wettable powder of niclosamide ethanolamine salt (WPNES) was prepared into solutions at a concentration of 2 g/L. The different concentrations of drugs were sprayed evenly, and the same amount of water was used as blank control. Snails were surveyed using the systematic sampling method 1, 3 and 7 days post-treatment, and snail survival was observed. A fish pond was selected in Nanjian County, and 2 kg 25% WPPS was evenly sprayed on the water surface to allow the effective concentration of 20 g/L. Fish mortality was estimated 8, 24, 48 and 72 h post-treatment. Results One-day treatment with 1 and 2 g/L WPPS and 2 g/L WPNES resulted in 97.99%, 97.99% and 94.11% adjusted snail mortality rates (χ2 = 3.509 and 3.509, both P values > 0.05), and the adjusted snail mortality was all 100% 3 d post-treatment with 1 and 2 g/L WPPS and 2 g/L WPNES, while 7-day treatment with 1 and 2 g/L WPPS and 2 g/L WPNES resulted in 91.75%, 86.57% and 57.76% adjusted snail mortality rates (χ2 = 14.893 and 42.284, both P values < 0.05). Treatment with 2 g/L WPPS for 72 h resulted in a 0.67% cumulative mortality rate of fish. Conclusion 25% WPPS is effective for snail control and highly safe for fish, which is feasible for use in hilly schistosomiasis-endemic regions.

Measurement of morphological features of Oncomelania hupensis shells in Yunnan Province / 中国血吸虫病防治杂志

Objective To investigate the morphological variation of Oncomelania hupensis shells in Yunnan Province, so as to provide insights into the understanding of O. hupensis genetic evolution and control. Methods According to the O. hupensis density, geographical location, altitude, water system and environmental type, 12 administrative villages were sampled from 10 schistosomiasis-endemic counties (districts) in 3 prefectures (cities) of Yunnan Province as snail collection sites. From December 2021 to January 2022, about 200 snails were collected from each collection site, among which thirty adult snails (6 to 7 spirals) were randomly selected from each site, and the 11 morphological indexes of snail shells were measured and subjected to cluster analysis and principal component analysis. Results Of O. hupensis snails from 12 localities of Yunnan Province, the longest shell (7.33 mm) was detected in snails from Yongle Village, Eryuan County, with the shortest (4.68 mm) in Dongyuan Village, Gucheng District, and the largest angle of apex (59.47°) was measured in snails from Caizhuang Village, Midu County, with the smallest (41.40°) in Qiandian Village, Eryuan County. The mean coefficient of variation was 9.075% among O. hupensis snails from 12 localities of Yunnan Province, with the largest coefficient of variation seen in the thickness of the labra brim (29.809%). Among O. hupensis snails from 12 localities of Yunnan Province, the mean Euclidean distance was 2.26, with the shortest Euclidean distance seen between O. hupensis snails from Qiandian Village of Eryuan County and Wuxing Village of Dali City (0.26), and the largest found between O. hupensis snails from Caizhuang Village of Midu County and Cangling Village of Chuxiong County (8.17). Cluster analysis and principal component analysis classified O. hupensis snails from 12 localities of Yunnan Province into three categories, including the O. hupensis snail samples from Caizhuang Village of Midu County, O. hupensis snail samples from Cangling Village of Chuxiong County, and O. hupensis snail samples from Qiandian Village of Eryuan County, Wuxing Village of Dali City, Yangwu Village of Yongsheng County, Xiaoqiao Village of Xiangyun County, Yongle Village of Eryuan County, Xiaocen Village of Dali City, Anding Village of Nanjian County, Dongyuan Village of Gucheng District, Lianyi Village of Heqing County, and Dianzhong Village of Weishan County. The variations in these three categories of snail samples were mainly measured in the principal component 2 related to the angle of apex and the thickness of the labra brim. Conclusions The variations in the Euclidean distance and morphological features of shells of O. hupensis from 12 localities of Yunnan Province gradually rise with the decrease in the latitude of the collection sites. The angle of apex is an indicator for the growth of O. hupensis whorl.

Detection of bloodworm larvae (Diptera: Chironomidae) in the golden apple snail Pomacea canaliculata (Lamarck, 1819) (Gastropoda: Ampullariidae) in Metro Manila

Background@#Pomacea canaliculata (Lamarck, 1819) is an invasive freshwater snail in the Philippines that damages crops but is consumed as food. It is known to harbor parasites, some of which are pathogenic to humans. @*Objectives@#The objective of this study is to examine P. canaliculata individuals present in Metro Manila for parasite infection and identify the parasites and other organisms associated with the snail using molecular identification. @*Methodology@#P. canaliculata were collected from rivers and marketplaces in Metro Manila. Individuals were crushed and digested in Ash's digestive fluid and observed under a microscope. Collected parasites were subjected to DNA barcoding of the COI gene for putative identification. @*Results@#A total of 462 snails were gathered from 15 sites, eight of which were market areas that sourced the snails from outside Metro Manila. No known parasites were found. Two snails were found to contain insect larvae in the mantle (0.43% infection). The closest BLAST matches for the two insect larvae were the chironomid fly Nilodorum tainanus (91.0% identity) from a snail in Sucat, Muntinlupa, and another chironomid Parachironomus sp. (92.8% identity) found in a snail originally from Cavite and brought to Calumpang, Marikina. @*Conclusion@#This study is the first report of the presence of chironomids in Philippine P. canaliculata. This could have an impact on the allergenic status of these mollusks if consumed while containing these chironomids. The absence of infection of other medically important parasites is possibly due to the patchy distribution of the snails and few interactions with the definitive hosts of known parasites.

Ethanol Extract of Yacon Leaves (Smallanthus sonchifolius) Attenuates Fibroblast and Myofibroblast Expansion in Association With Downregulation of TGF β1 and Snail mRNA Expression in 5/6-Subtotal Nephrectomy Model in Mice

@#Introduction: Chronic kidney disease (CKD) is characterized by fibroblast activation, myofibroblast formation, and up-regulation of transforming growth factor-β1 (TGF-β1) that may activate Snail in fibroblast to myofibroblast transition. Ethanol extract of Yacon leaves is known to have a renoprotective effect on diabetic nephropathy but its effect in the CKD model is unknown. This experimental study aimed to elucidate the effect of ethanol extract from Yacon leaves in attenuating renal failure in a CKD mice model. Methods: Male Swiss-Webster mice (3 months, 30–40 grams, n=25) underwent 5/6 subtotal nephrectomy (SN) to induce CKD. The mice were divided into five groups: SN, SN mice with oral treatment of Yacon leaves ethanol extract with doses 0.735 μg/kg (SN+YK1), 1.47 μg/kg (SN+YK2), and 2.94 μg/kg (SN+YK3), and a Sham operation (SO) group with aquadest 0.1% supplementation. Mice were euthanized on day 14 after the operation and kidneys were harvested. Paraffin sections were used for histological analysis. Immunostaining was done for quantifying fibroblasts and myofibroblasts. We performed RT-PCR to measure TGF-β1 and Snail mRNA expressions. Results: The SN group had significantly higher fibroblast number, myofibroblast fraction area, TGF-β1 and Snail mRNA expressions compared to the SO. The fibroblasts number (p<0.001) and myofibroblast fraction areas (p<0.001) were significantly lower in Yacon treated-groups compared to the SN group. RT-PCR analysis showed lower mRNA expressions of TGF-β1 and Snail, but no significant differences were found among the various Yacon treated-groups. Conclusion: Ethanol extracts of Yacon leaves improved kidney damage in male mice with 5/6 subtotal nephrectomy model.