Meta-analysis contrasting freshwater biodiversity in forests and oil palm plantations with and without riparian buffers.

The expansion of oil palm plantations has led to land-use change and deforestation in the tropics, which has affected biodiversity. Although the impacts of the crop on terrestrial biodiversity have been extensively reviewed, its effects on freshwater biodiversity remain relatively unexplored. We reviewed the research assessing the impacts of forest-to-oil palm conversion on freshwater biota and the mitigating effect of riparian buffers on these impacts. We searched for studies comparing taxa richness, species abundance, and community composition of macroinvertebrates, amphibians, and fish in streams in forests (primary and disturbed) and oil palm plantations with and without riparian buffers. Then, we conducted a meta-analysis to quantify the overall effect of the land-use change on the 3 taxonomic groups. Twenty-nine studies fulfilled the inclusion criteria. On average, plantations lacking buffers hosted 44% and 19% fewer stream taxa than primary and disturbed forests, respectively. Stream taxa on plantations with buffers were 24% lower than in primary forest and did not differ significantly from disturbed forest. In contrast, stream community composition differed between forests and plantations regardless of the presence of riparian buffers. These differences were attributed to agrochemical use and altered environmental conditions in the plantations, including temperature changes, worsened water conditions, microhabitat loss, and food and shelter depletion. On aggregate, abundance did not differ significantly among land uses because increases in generalist species offset the population decline of vulnerable forest specialists in the plantation. Our results reveal significant impacts of forest-to-oil palm conversion on freshwater biota, particularly taxa richness and composition (but not aggregate abundance). Although preserving riparian buffers in the plantations can mitigate the loss of various aquatic species, it cannot conserve primary forest communities. Therefore, safeguarding primary forests from the oil palm expansion is crucial, and further research is needed to address riparian buffers as a promising mitigation strategy in agricultural areas.

Metaanálisis contrastando la biodiversidad de agua dulce en los bosques y las plantaciones de palma de aceite con o sin bosques ribereños Resumen La expansión de las plantaciones de palma de aceite ha derivado en cambios en el uso de suelo y deforestación en los trópicos, afectando a la biodiversidad. Existe una revisión extensa del impacto de este cultivo sobre la biodiversidad terrestre, pero sus efectos sobre la biodiversidad de agua dulce todavía no están muy documentados. Revisamos las investigaciones que han evaluado el impacto de la conversión de bosque a plantación de palma de aceite sobre la biota de agua dulce y el efecto mitigante que tienen los bosques ribereños sobre este impacto. Buscamos estudios que compararan la riqueza de taxones, abundancia de especies y composición comunitaria de los macroinvertebrados, anfibios y peces en los riachuelos de los bosques (primarios y perturbados) y los sembradíos de palma de aceite con y sin bosques ribereños. Después realizamos un metaanálisis para cuantificar el efecto del cambio de uso de suelo en los tres grupos taxonómicos. Veintinueve estudios cumplieron con el criterio de inclusión. En promedio, las plantaciones carentes de bosques ribereños albergaron 44% y 19% menos taxones que los bosques primarios y perturbados. Los taxones en los sembradíos con bosques ribereños fueron 24% menos que en el bosque primario y no difirieron significativamente del bosque perturbado. Como contraste, la composición comunitaria del riachuelo difirió entre los bosques y los sembradíos sin importar la presencia de los bosques ribereños. Atribuimos estas diferencias al uso de agroquímicos y las condiciones ambientales alteradas en las plantaciones, incluidas los cambios térmicos, condiciones hidrológicas alteradas, pérdida de microhábitats y reducción de alimentos y refugios. En general, la abundancia no difirió significativamente entre los usos de suelo porque el incremento de especies generalistas en las plantaciones contrarresta la declinación poblacional de los especialistas de bosque vulnerables. Nuestros resultados revelan un impacto significativo de la conversión de bosque a plantación sobre la biota de agua dulce, particularmente la riqueza de taxones y la composición (pero no la abundancia agregada). Aunque mantener los bosques ribereños en las plantaciones puede mitigar la pérdida de varias especies acuáticas, no puede conservar las comunidades del bosque primario. Por lo tanto, es crucial salvaguardar los bosques primarios de la expansión del aceite de palma, además de que se necesitan más investigaciones para abordar los bosques ribereños como una estrategia prometedora de mitigación en las áreas agrícolas.

Effect of an Oral Bivalent Vaccine on Immune Response and Immune Gene Profiling in Vaccinated Red Tilapia (Oreochromis spp.) during Infections with Streptococcus iniae and Aeromonas hydrophila.

Streptococcosis and aeromonasis inflicted by Streptococcus iniae and Aeromonas hydrophila, respectively, have affected tilapia industries worldwide. In this study, we investigated antibody responses and explored the mechanisms of protection rendered by an oral bivalent vaccine in red tilapia following challenges with S. iniae and A. hydrophila. The results of specific IgM antibody response revealed that the IgM titers against S. iniae and A. hydrophila in the bivalent incorporated (BI) vaccine group were significantly higher (p < 0.05) than those in the bivalent spray (BS) vaccine fish and unvaccinated control fish throughout the experiment. Real-time qPCR results also showed that the gene expression of CD4, MHC-I, MHC-II, IgT, C-type lysozyme, IL-1ß, TNF-α, and TGF-ß remained significantly higher (p < 0.05) than that of the controls between 24 and 72 h post-infection (hpi) in both mucosal (hindgut) and systemic (spleen and head−kidney) organs of BI vaccinated fish. Furthermore, the highest relative expression of the TGF-ß, C-type lysozyme, and IgT genes in the BI vaccinated group was observed in the challenged fish's spleen (8.8-fold), head kidney (4.4-fold), and hindgut (19.7-fold) tissues, respectively. The present study suggests that the bivalent incorporated (BI) vaccine could effectively improve the immune function and activate both humoral and cell-mediated immunities in vaccinated red tilapia following the bacterial challenges.

Prevalence, Antibiotics Resistance and Plasmid Profiling of Vibrio spp. Isolated from Cultured Shrimp in Peninsular Malaysia.

Vibrio is the most common bacterium associated with diseases in crustaceans. Outbreaks of vibriosis pose a serious threat to shrimp production. Therefore, antibiotics are commonly used as preventative and therapeutic measures. Unfortunately, improper use of antibiotics leads to antibiotic resistance. Nevertheless, information on the occurrence of Vibrio spp. and antibiotic use in shrimp, particularly in Malaysia, is minimal. This study aimed to provide information on the occurrence of Vibrio spp., its status of antibiotic resistance and the plasmid profiles of Vibrio spp. isolated from cultured shrimp in Peninsular Malaysia. Shrimp were sampled from seven farms that were located in different geographical regions of Peninsular Malaysia. According to the observations, 85% of the shrimp were healthy, whereas 15% were unhealthy. Subsequently, 225 presumptive Vibrio isolates were subjected to biochemical tests and molecular detection using the pyrH gene. The isolates were also tested for antibiotic susceptibility against 16 antibiotics and were subjected to plasmid profiling. Eventually, 13 different Vibrio spp. were successfully isolated and characterized using the pyrH gene. They were the following: V. parahaemolyticus (55%), V. communis (9%), V. campbellii (8%), V. owensii (7%), V. rotiferianus (5%), Vibrio spp. (4%), V. alginolyticus (3%), V. brasiliensis (2%), V. natriegens (2%), V. xuii (1%), V. harveyi (1%), V. hepatarius (0.4%) and P. damselae (3%). Antibiotic susceptibility profiles revealed that all isolates were resistant to penicillin G (100%), but susceptible to norfloxacin (96%). Furthermore, 16% of the isolates revealed MAR of less than 0.2, while 84% were greater than 0.2. A total of 125 isolates harbored plasmids with molecular weights between 1.0 and above 10 kb, detected among the resistant isolates. The resistant isolates were mediated by both chromosomal and plasmid factors. These findings support the use of surveillance data on the emerging patterns of antimicrobial-resistance and plasmid profiles of Vibrio spp. in shrimp farms. The findings from this study can be used to develop a better disease management strategy for shrimp farming.

Draft genome sequence of myo-inositol utilizing Aeromonas dhakensis 1P11S3 isolated from striped catfish (Pangasianodon hypopthalmus) in a local fish farm in Malaysia.

A hypervirulent pathotype of A. hydrophila (vAh) is responsible for Motile Aeromonas Septicemia (MAS) and causes mass mortalities among farmed carp and catfish species in the USA and China. One unique phenotype for vAh among other A. hydrophila strains is the ability to utilize myo-inositol as a sole carbon source. While screening for Aeromonas isolates from diseased fish that can grow using myo-inositol as a sole carbon source, A. dhakensis 1P11S3 was isolated from the spleen of striped catfish (Pangasianodon hypopthalmus) displaying clinical MAS symptoms from a freshwater farm in Malaysia. Aeromonas dhakensis is also an important pathogen in aquaculture, and in this study, we report the draft genome sequence for A. dhakensis 1P11S3, that utilize myo-inositol as a sole carbon source.

Laboratory and Field Assessments of Oral Vibrio Vaccine Indicate the Potential for Protection against Vibriosis in Cultured Marine Fishes.

Vibriosis is one of the most common threats to farmed grouper; thus, substantial efforts are underway to control the disease. This study presents an oral vaccination against multiple Vibrio spp. in a marine fish with double booster immunisation. The Vibrio harveyi strain VH1 vaccine candidate was selected from infected groupers Epinephelus sp. in a local farm and was formalin inactivated and combined with commercial feed at a 10% ratio (v/w). A laboratory vaccination trial was conducted for seventy days. The induction of IgM antibody responses in the serum of Asian seabass Lates calcarifer immunised with the oral Vibrio harveyi strain VH1 was significantly (p < 0.05) increased as early as week one post-primary vaccination. Subsequent administration of the first and second booster for 5 consecutive days, starting on days 14 and 42, respectively, improved the specific antibody level and reached a highly significant (p < 0.05) value at days 35 and 49 before slightly decreasing from day 56 onwards. Antibody titres of the control unvaccinated group remained relatively stable and low throughout the experimental period. At the end of the 70-day vaccination trial, 23 days post final boost, an intraperitoneal challenge with a field strain of Vibrio harveyi, V. alginolyticus, and V. parahaemolyticus was carried out. Our challenge study showed that oral Vibrio harveyi strain VH1 vaccine candidate could induce significant protection, with an RPS of 70-80% against different Vibrio species. Thereafter, a field trial was conducted in a mariculture farm to study the effect of field vaccination using the oral Vibrio harveyi strain VH1 vaccine candidate. A total of 3000 hybrid grouper juveniles were divided into two groups in triplicate. Fish of Group 1 were not vaccinated, while Group 2 were vaccinated with the feed-based vaccine. Vaccinations were carried out on days 0, 14, and 42 via feeding the fish with the vaccine at 4% body weight for 5 consecutive days. At the end of the study period, the fish survival rate was 80% for the vaccinated group, significantly (p < 0.05) higher than the 65% seen in the control unvaccinated group. Furthermore, the vaccinated fish showed significantly (p < 0.05) better growth performances. Therefore, the oral Vibrio vaccine from the inactivated Vibrio harveyi strain VH1 is a potential versatile vaccine candidate that could stimulate good immune responses and confer high protection in both Asian seabass, Lates calcarifer, and farm hybrid grouper Epinephelus fuscoguttatus × Epinephelus lanceolatus.

Field Efficacy of a Feed-Based Inactivated Vaccine against Vibriosis in Cage-Cultured Asian Seabass, Lates calcarifer, in Malaysia.

Vibrio spp. are important aquaculture pathogens that cause vibriosis, affecting large numbers of marine fish species. This study determines the field efficacy of a feed-based inactivated vaccine against vibriosis in cage-cultured Asian seabass. A total of 4800 Asian seabass, kept in a field environment, were separated equally into two groups (vaccinated and non-vaccinated) in duplicate. Fish of Group 1 were orally administered the feed-based vaccine on weeks 0 (prime vaccination), 2 (booster), and 6 (second booster) at 4% body weight, while the non-vaccinated fish of Group 2 were fed with a commercial formulated pellet without the vaccine. Fish gut, mucus, and serum were collected, the length and weight of the fish were noted, while the mortality was recorded at 2-week intervals for a period of 16 weeks. The non-specific lysozyme activities were significantly (p < 0.05) higher in the fish of Group 1 than the non-vaccinated fish of Group 2. Similarly, the specific IgM antibody levels in serum and mucus were significantly (p < 0.05) higher in Group 1 than in Group 2, as seen in the second week, with the highest level 8 weeks after primary immunization. At week 16, the growth performance was significantly (p < 0.05) better in Group 1 and showed lower bacterial isolation in the gut than Group 2. Despite the statistical insignificance (p > 0.05), the survival rate was slightly higher in Group 1 (71.3%) than Group 2 (67.7%). This study revealed that feed-based vaccination improves growth performance, stimulates innate and adaptive immune responses, and increases protection of cultured Asian seabass, L. calcarifer, against vibriosis.

Comparative Pathogenicity of Aeromonas spp. in Cultured Red Hybrid Tilapia (Oreochromis niloticus × O. mossambicus).

The genus Aeromonas has been recognised as an important pathogenic species in aquaculture that causes motile Aeromonas septicaemia (MAS) or less severe, chronic infections. This study compares the pathogenicity of the different Aeromonas spp. that were previously isolated from freshwater fish with signs of MAS. A total of 124 isolates of Aeromonas spp. were initially screened for the ability to grow on M9 agar with myo-inositol as a sole carbon source, which is a discriminatory phenotype for the hypervirulent A. hydrophila (vAh) pathotype. Subsequently, LD50 of six selected Aeromonas spp. were determined by intraperitoneal injection of bacterial suspension containing 103, 105, and 107 CFU/mL of the respective Aeromonas sp. to red hybrid tilapias. The kidneys, livers and spleens of infected moribund fish were examined for histopathological changes. The screening revealed that only A. dhakensis 1P11S3 was able to grow using myo-inositol as a sole carbon source, and no vAh strains were identified. The LD50-240h of A. dhakensis 1P11S3 was 107 CFU/mL, while the non-myo-inositol utilizing A. dhakensis 4PS2 and A. hydrophila 8TK3 was lower at 105 CFU/mL. Similarly, tilapia challenged with the myo-inositol A. dhakensis 1P11S3 showed significantly (p < 0.05) less severe signs, gross and histopathological lesions, and a lower mortality rate than the non-myo-inositol A. dhakensis 4PS2 and A. hydrophila 8TK3. These findings suggested that myo-inositol utilizing A. dhakensis 1P11S3 was not a hypervirulent Aeromonas sp. under current experimental disease challenge conditions, and that diverse Aeromonas spp. are of concern in aquaculture farmed freshwater fish. Therefore, future study is warranted on genomic level to further elucidate the influence of myo-inositol utilizing ability on the pathogenesis of Aeromonas spp., since this ability correlates with hypervirulence in A. hydrophila strains.

Invasion Risk and Potential Impact of Alien Freshwater Fishes on Native Counterparts in Klang Valley, Malaysia.

This study explores the potential effects of alien fishes on the native fish community, well-being, and tropic preferences in selected rivers of Klang Valley, Malaysia. Following the Aquatic Species Invasiveness Screening Kit assessment, most of the alien fishes (80%) are invasive. The alien species occurrences correlated positively (p < 0.05) with poor water quality, such as rivers with high ammonia-nitrogen and nitrite, but negatively with phosphate and dissolved oxygen. Anthropogenic characteristics, such as rivers with high pollution levels and ease of accessibility to the fish habitat, are mainly associated positively (p < 0.05) with the occurrences of alien fish species. In general, the results of fish stomach contents analyses and their associated indices, together with stable carbon and nitrogen isotopes, revealed domination by alien fishes or diet overlaps between both alien and native fish species. This finding indicates that alien fishes benefited from the impacts of the anthropogenic activities in their surrounding habitats, while their plasticity in feeding habits might help them to invade, survive, and dominate in the rivers of Klang Valley, Malaysia.

Polystyrene Microplastics Exposure: An Insight into Multiple Organ Histological Alterations, Oxidative Stress and Neurotoxicity in Javanese Medaka Fish (Oryzias javanicus Bleeker, 1854).

Microplastics (MPs) have become pollutants of concern due to their unknown human health effect and negative impact on terrestrial and aquatic ecosystems. There is increasing number of experimental research on MPs globally with its effects not fully understood; recent animal studies explore its effects on the intestines, yet on other vital organs. Javanese medaka fish was exposed to polystyrene microplastics (PS-MPs) beads for a period of 21 days. Histological alterations, intestinal oxidative stress, permeability and neurotoxicity were evaluated. Significant inflammatory changes and tissue damage were observed in the intestine, liver and kidney. Intestinal oxidative stress and permeability were found to be significantly increased. In the brain, neurotoxicity characterised by a significant induction of oxidative stress, lipid peroxidation and the inhibition of acetylcholinesterase enzyme were elucidated. This study provided an insight into the multiple organ effect of microplastics exposure, necessitating further exploration and identification of biomarkers to be utilised for biomonitoring population at risk in the future.

Vaccine Efficacy of a Newly Developed Feed-Based Whole-Cell Polyvalent Vaccine against Vibriosis, Streptococcosis and Motile Aeromonad Septicemia in Asian Seabass, Lates calcarifer.

Multiple infections of several bacterial species are often observed under natural farm conditions. The infections would cause a much more significant loss compared to a single infectious agent. Vaccination is an essential strategy to prevent diseases in aquaculture, and oral vaccination has been proposed as a promising technique since it requires no handling of the fish and is easy to perform. This research attempts to develop and evaluate a potential feed-based polyvalent vaccine that can be used to treat multiple infections by Vibrios spp., Streptococcus agalactiae, and Aeromonas hydrophila, simultaneously. The oral polyvalent vaccine was prepared by mixing formalin-killed vaccine of V. harveyi, S. agalactiae, and A. hydrophila strains with commercial feed pellet, and palm oil as an adjuvant was added to improve their antigenicity. Thereafter, a vaccinated feed pellet was tested for feed quality analysis in terms of feed stability in water, proximate nutrient analysis, and palatability, safety, and growth performance using Asian seabass, Lates calcarifer as a fish host model. For immune response analysis, a total of 300 Asian seabass juveniles (15.8 ± 2.6 g) were divided into two groups in triplicate. Fish of group 1 were not vaccinated, while group 2 was vaccinated with the feed-based polyvalent vaccine. Vaccinations were carried out on days 0 and 14 with oral administration of the feed containing the bacterin at 5% body weight. Samples of serum for antibody and lysozyme study and the spleen and gut for gene expression analysis were collected at 7-day intervals for 6 weeks. Its efficacy in protecting fish was evaluated in aquarium challenge. Following vaccination by the polyvalent feed-based vaccine, IgM antibody levels showed a significant (p < 0.05) increase in serum against Vibrio harveyi, Aeromonas hydrophila, and Streptococcus agalactiae and reached the peak at week 3, 5, and 6, respectively. The high-stimulated antibody in the serum remained significantly higher than the control (p < 0.05) at the end of the 6 weeks vaccination trial. Not only that, but the serum lysozyme level was also increased significantly at week 4 (p < 0.05) as compared to the control treatment. The immune-related gene, dendritic cells, C3, Chemokine ligand 4 (CCL4), and major histocompatibility complex class I (MHC I) showed significantly higher expression (p < 0.05) after the fish were vaccinated with the oral vaccine. In the aquarium challenge, the vaccine provided a relative percentage survival of 75 ± 7.1%, 80 ± 0.0%, and 80 ± 0.0% after challenge with V. harveyi, A. hydrophila, and S. agalactiae, respectively. Combining our results demonstrate that the feed-based polyvalent vaccine could elicit significant innate and adaptive immunological responses, and this offers an opportunity for a comprehensive immunization against vibriosis, streptococcosis, and motile aeromonad septicemia in Asian seabass, Lates calcarifer. Nevertheless, this newly developed feed-based polyvalent vaccination can be a promising technique for effective and large-scale fish immunization in the aquaculture industry shortly.

Immuno-protective efficiency of feed-based whole-cell inactivated bivalent vaccine against Streptococcus and Aeromonas infections in red hybrid tilapia (Oreochromis niloticus × Oreochromis mossambicus).

Streptococcosis and motile aeromonad septicemia (MAS) are well-known diseases in tilapia culture, which cause mass mortality with significant economic losses. The development of feed-based bivalent vaccines in controlling these diseases has been initiated, however, the mechanisms of immunities and cross-protection in fish remain unclear. This study was conducted to assess the immuno-protective as well as the cross-protective efficacy of a newly developed feed-based bivalent vaccine against Streptococcus and Aeromonas infections in red hybrid tilapia. A total of five groups of fish were vaccinated orally through two different techniques; bivalent vaccine (inactivated Streptococcus iniae and Aeromonas hydrophila) sprayed on feed pellets (BS group); bivalent vaccine (inactivated S. iniae and A. hydrophila) incorporated in feed (BI group); monovalent inactivated S. iniae and A. hydrophila vaccine separately incorporated into feed as monovalent S. iniae (MS group) and monovalent A. hydrophila (MA group); and control group (without vaccine). The feed-based vaccine was delivered orally at 5% of body weight for five consecutive days. The booster doses were given in the same manner on weeks 2 and 6. Serum and skin mucus samples were collected to assess the IgM responses using indirect ELISA. The first administration of the feed-based vaccine stimulated the IgM levels that lasted until week 3, while the second booster ensured that the IgM levels remained high for a period of 16 weeks in the BI, MS and MA groups. The BI group developed a strong and significantly (P < 0.05) higher systemic and mucosal IgM responses against both S. iniae and A. hydrophila and also cross-protective antigen S. agalactiae and A. veronii compared to the BS and control groups. Quantitative real-time PCR results also showed that the relative expressions of IL-8, INF-γ and IgM in the BI immunized fish spleen, head kidney and hindgut exhibited various significant (P < 0.05) rising trends following both the vaccination and the challenge phase. On weeks 10, all fish were challenged through the intraperitoneal route, where relative percent survivals (RPS) of 82.22 ± 3.85% when challenged with S. iniae, 77.78 ± 3.85% when challenged with A. hydrophila and 77.78 ± 3.85% when co-challenged with both S. iniae and A. hydrophila were observed in the BI group, which were significantly higher (P < 0.05) compared to the other groups. The BI group also showed significantly (P < 0.05) higher partial cross-protections following challenges with S. agalactiae (RPS at 60.00 ± 6.67%) and A. veronii (RPS at 57.78 ± 7.70%). This study demonstrated that immunization with feed-based BI vaccine elicited immune responses that were capable of protecting red hybrid tilapia against streptococcosis and MAS.

Microplastics Pollution as an Invisible Potential Threat to Food Safety and Security, Policy Challenges and the Way Forward.

Technological advances, coupled with increasing demands by consumers, have led to a drastic increase in plastic production. After serving their purposes, these plastics reach our water bodies as their destination and become ingested by aquatic organisms. This ubiquitous phenomenon has exposed humans to microplastics mostly through the consumption of sea food. This has led the World Health Organization (WHO) to make an urgent call for the assessment of environmental pollution due to microplastics and its effect on human health. This review summarizes studies between 1999 and 2020 in relation to microplastics in aquatic ecosystems and human food products, their potential toxic effects as elicited in animal studies, and policies on their use and disposal. There is a paucity of information on the toxicity mechanisms of microplastics in animal studies, and despite their documented presence in food products, no policy has been in place so far, to monitor and regulates microplastics in commercial foods meant for human consumption. Although there are policies and regulations with respect to plastics, these are only in a few countries and in most instances are not fully implemented due to socioeconomic reasons, so they do not address the problem across the entire life cycle of plastics from production to disposal. More animal research to elucidate pathways and early biomarkers of microplastic toxicity that can easily be detected in humans is needed. This is to create awareness and influence policies that will address this neglected threat to food safety and security.

Co-Infections of Tilapia Lake Virus, Aeromonas hydrophila and Streptococcus agalactiae in Farmed Red Hybrid Tilapia.

A high death rate among red hybrid tilapias was observed in a farm in Selangor, Malaysia, in January 2020. The affected fish appeared lethargic, isolated from schooling group, showed loss of appetite, red and haemorrhagic skin, exophthalmia and enlarged gall bladders. Histopathological assessment revealed deformation of kidney tubules, and severe congestion with infiltrations of inflammatory cells in the brains and kidneys. Syncytial cells and intracytoplasmic inclusion bodies were occasionally observed in the liver and brain sections. Tilapia Lake Virus (TiLV), Aeromonas hydrophila and Streptococcus agalactiae were identified in the affected fish, either through isolation or through PCR and sequencing analysis. The phylogenetic tree analysis revealed that the TiLV strain in this study was closely related to the previously reported Malaysian strain that was isolated in 2019. On the other hand, A. hydrophila and S. agalactiae were closer to Algerian and Brazilian strains, respectively. The multiple antibiotic resistance index for A. hydrophila and S. agalactiae was 0.50 and 0.25, respectively. Co-infections of virus and bacteria in cultured tilapia is a new threat for the tilapia industry.

Species composition and invasion risks of alien ornamental freshwater fishes from pet stores in Klang Valley, Malaysia.

The ornamental fish trade has been considered as one of the most important routes of invasive alien fish introduction into native freshwater ecosystems. Therefore, the species composition and invasion risks of fish species from 60 freshwater fish pet stores in Klang Valley, Malaysia were studied. A checklist of taxa belonging to 18 orders, 53 families, and 251 species of alien fishes was documented. Fish Invasiveness Screening Test (FIST) showed that seven (30.43%), eight (34.78%) and eight (34.78%) species were considered to be high, medium and low invasion risks, respectively. After the calibration of the Fish Invasiveness Screening Kit (FISK) v2 using the Receiver Operating Characteristics, a threshold value of 17 for distinguishing between invasive and non-invasive fishes was identified. As a result, nine species (39.13%) were of high invasion risk. In this study, we found that non-native fishes dominated (85.66%) the freshwater ornamental trade in Klang Valley, while FISK is a more robust tool in assessing the risk of invasion, and for the most part, its outcome was commensurate with FIST. This study, for the first time, revealed the number of high-risk ornamental fish species that give an awareness of possible future invasion if unmonitored in Klang Valley, Malaysia.

Multilocus Sequence Typing Analysis of Invasive and Non-Invasive Group B Streptococcus of Hospital Origin in Malaysia.

The aim of this study was to study the genotype of a hospital collection of Group B Streptococcus (GBS) from invasive and non-invasive sites. Fifty-one pre-characterised human of GBS were re-identified and further analysed by multilocus sequence typing (MLST) in relation to previously published serotypes. Fifteen sequence types (ST) were found with ST1 being the most predominant. ST1 was also associated with majority of the invasive isolates. The genotypic distribution patterns of GBS in this study were largely in agreement with previous reports from other countries indicating the tendency of certain genotypes to prevail in human infection settings.

Prevalence, antimicrobial susceptibility and plasmid profiling of Vibrio spp. isolated from cultured groupers in Peninsular Malaysia.

BACKGROUND: Numerous prevalence studies of Vibrio spp. infection in fish have been extensively reported worldwide, including Malaysia. Unfortunately, information on the prevalence of Vibrio spp. in groupers (Epinephelus spp.) is limited. In this study, groupers obtained from nine farms located at different geographical regions in Malaysia were sampled for the presence of pathogenic Vibrio spp. and their susceptibility profiles against seven antibiotics. RESULTS: Out of 270 grouper samples, 195 (72%) were detected with the presence of Vibrio spp. Vibrio communis showed highest prevalence in grouper (28%), followed by V. parahaemolyticus (25%), V. alginolyticus (19%), V. vulnificus (14%), V. rotiferianus (3%), Vibrio sp. (3%), V. campbellii (2%), V. mytili (2%), V. furnissii (2%), V. harveyi (1%), V. tubiashii (1%), V. fluvialis (0.3%) and V. diabolicus (0.3%). Assessment on the antibiotic susceptibility profiles of the Vibrio spp. revealed that majority of the isolates were susceptible to tetracycline, streptomycin, erythromycin and bacitracin, but resistance to ampicillin, penicillin G and vancomycin. The mean MAR index of the Vibrio isolates was 0.51, with 85% of the isolates showed MAR index value of higher than 0.2. Results indicate that the Vibrio spp. were continuously exposed to antibiotics. Furthermore, the plasmid profiles of Vibrio spp. showed that 38.7% of the isolates harbored plasmid with molecular weight of more than 10 kb, while 61.3% were without plasmid. During curing process, Vibrio spp. lost their plasmid, but remained resistant to ampicillin, penicillin G, bacitracin and vancomycin while a few isolates remained resistant to erythromycin, streptomycin and tetracycline. The results suggested that the resistance to antibiotics in isolated Vibrio spp. might be due to chromosomal and plasmid borne. CONCLUSIONS: This study demonstrates the prevalence of Vibrio spp. in groupers and the distribution of multidrug resistance strains that could be of concern to the farmers in Malaysia. In addition, data from this study can be further used in fish disease management plan.

Virulence-associated genes and antibiotic resistance patterns of Vibrio spp. isolated from cultured marine fishes in Malaysia.

BACKGROUND: Vibriosis is an important bacterial disease of cultured marine fishes worldwide. However, information on the virulence and antibiotic resistance of Vibrio spp. isolated from fish are scarce. This study investigates the distribution of virulence associated genes and antibiotic resistance patterns of Vibrio spp. isolated from cage-cultured marine fishes in Malaysia. RESULTS: A total of 63 Vibrio spp. isolated from 62 cultured marine fishes in various geographical regions in Peninsular Malaysia were analysed. Forty-two of the isolates (66.7%) were positive for all chiA, luxR and vhpA, the virulence genes produced by pathogenic V. harveyi. A total of 62 Vibrio isolates (98%) had tlh gene of V. parahaemolyticus, while flaC gene of V. anguillarum was detected in 43 of isolates (68%). Other virulence genes, including tdh, trh, hlyA and toxRvc were absent from any of the isolates. Multiple antibiotic resistance (MAR) was exhibited in all strains of Harveyi clade, particularly against ampicillin, penicillin, polypeptides, cephems and streptomycin. The MAR index ranged between 0.06 and 0.56, and 75% of the isolates have MAR index of higher than 0.20. Host species and geographical origin showed no correlation with the presence of virulence genes and the antibiotic resistance patterns of Vibrio spp. CONCLUSIONS: The study indicates that majority of Vibrio spp. isolated from cultured marine fishes possess virulence genes, but were not associated with human pathogen. However, the antibiotics resistance is a real concern and warrants ongoing surveillance. These findings represent an updated knowledge on the risk of Vibrio spp. to human health, and also provides valuable insight on alternative approaches to combat vibriosis in cultured fish.

Study on Streptococcus agalactiae infection in Javanese medaka (Oryzias javanicus Bleeker, 1854) model.

This study determines the median lethal dose, and describes the clinico-pathological changes and disease development following Streptococcus agalactiae infection in Javanese medaka model. Javanese medakas were infected with S. agalactiae via intraperitoneal (IP) from 104 to 108 CFU/mL, and immersion (IM) route from 103 to 107 CFU/mL. The LD50-240h and clinico-pathological changes of the fish was determined until 240 h post infection (hpi). Next, the disease development was determined for 96 hpi in the fish following IP and IM infection at 103 CFU/mL and 104 CFU/mL, respectively. The LD50-240h of S. agalactiae in Javanese medaka was lower following IP injection (4.5 × 102 CFU/mL), compared to IM route (3.5 × 103 CFU/mL). The clinical signs included separating from the schooling group, swimming at the surface of water column, lethargy, erratic swimming pattern, corneal opacity and exophthalmia. Histopathological examinations revealed generalized congestion in almost all internal organs, particularly in liver and brain, while the kidney displayed tubular necrosis. Both IP and IM routes showed significant positive correlation (p < 0.05) between the CFU/g of S. agalactiae in the fish tissue and fish deaths. Moreover, the lesions for histopathological scoring in selected organs following IP and IM challenges were also reflecting the CFU/g and fish deaths. This study indicates the capability of Javanese medaka as a model organism in study of streptococcosis development.

Environmental Factors Associated with the Presence of Vibrionaceae in Tropical Cage-Cultured Marine Fishes.

This study investigated the environmental factors associated with the presence of Vibrionaceae in economically important cage-cultured tropical marine fishes: the Asian Seabass Lates calcarifer, snapper Lutjanus sp., and hybrid grouper Epinephelus sp. Fish sampling was conducted at monthly intervals between December 2016 and August 2017. The body weight and length of individual fish were measured, and the skin, eye, liver, and kidney were sampled for bacterial isolation and identification. Water physicochemical parameters during the sampling activities were determined, and the enumeration of total Vibrionaceae count was also conducted from water and sediment samples. Nine species of Vibrio were identified, including V. alginolyticus, V. diabolicus, V. harveyi, V. campbellii, V. parahaemolyticus, V. rotiferianus, V. furnissii, V. fluvialis, and V. vulnificus. Photobacterium damselae subsp. damselae was also identified. A total of 73% of the isolated Vibrio belonged to the Harveyi clade, followed by the Vulnificus clade (5.5%) and Cholera clade (0.6%). Highest occurrence of Vibrio spp. and P. damselae subsp. damselae was found in hybrid grouper (72%), followed by Asian Seabass (48%) and snapper (36%). The associations of Vibrio spp. and P. damselae subsp. damselae with the host fish were not species specific. However, fish mortality and fish size showed strong associations with the presence of some Vibrio spp. On average, 60% of the infected cultured fish exhibited at least one clinical sign. Nevertheless, inconsistent associations were observed between the pathogens and water quality. The yearlong occurrence and abundance of Vibrionaceae in the environmental components indicate that they might serve as reservoirs of these pathogens.

Evaluation of the neurotoxic effects of chronic embryonic exposure with inorganic mercury on motor and anxiety-like responses in zebrafish (Danio rerio) larvae.

Chronic exposure to mercury (Hg) can lead to cumulative impairments in motor and cognitive functions including alteration in anxiety responses. Although several risk factors have been identified in recent year, little is known about the environmental factors that either due exposure toward low level of inorganic mercury that may led to the developmental disorders. The present study investigated the effects of embryonic exposure of mercury chloride on motor function and anxiety-like behavior. The embryo exposed to 6 different concentrations of HgCl2 (7.5, 15, 30, 100, 125, 250nM) at 5hpf until hatching (72hpf) in a semi-static condition. The mortality rate increased in a dose dependent manner where the chronic embryonic exposure to 100nM decreased the number of tail coiling, heartbeat, and swimming activity. Aversive stimulus was used to examine the effects of 100nM interferes with the development of anxiety-related behavior. No elevation in both thigmotaxis and avoidance response of 6dpf larvae exposed with 100nM were found. Biochemical analysis showed HgCl2 exposure affects proteins, lipids, carbohydrates and nucleic acids of the zebrafish larvae. These results showed that implication of HgCl2 on locomotor and biochemical defects affects motor performance and anxiety-like responses. Yet, the potential underlying mechanisms these responses need to be further investigated which is crucial to prevent potential hazards on the developing organism due to neurotoxicant exposure.