Effects on survival and bacterial community composition of the aquaculture water and gastrointestinal tract of shrimp (Litopenaeus vannamei) exposed to probiotic treatments after an induced infection of acute hepatopancreatic necrosis disease

Acute hepatopancreatic necrosis disease (AHPND) is a devastating condition impacting marine shrimp production worldwide. The objective of this study was to evaluate the effects of four probiotic formulations on Pacific white shrimp (Litopenaeus vannamei) infected with pathogenic Vibrio parahaemolyticus causing AHPND. In addition, bacterial community composition analyses of shrimp gastrointestinal tract (GIT) and aquaculture water before and after infection were conducted by sequencing variable region 4 of the 16S rRNA gene on the Illumina MiSeq platform. Treatments included: (1) Lactobacillus casei (P1), (2) L. casei and Rhodopseudomonas palustris (P2), (3) L. casei, Saccharomyces cerevisiae and R. palustris (P3), and (4) a commercial probiotic EM^® (EM), which showed shrimp survival of 11.7%, 26.7%, 36.7% and 73.3% respectively. Treatments causing lower survival showed greater relative abundance (>60%) of family Vibrionaceae in the GIT compared to treatments with higher survival. Diversity indices from GIT samples revealed that treatments showing higher survival had higher Shannon index values (4.69 ± 0.133), compared with those of treatments with lower survival (0.17 ± 0.004). Diversity indices from water samples did not show significant differences after infection (Shannon index 4.64 ± 0.53). The results showed that probiotics could effectively mitigate AHPND while maintaining diverse microbial composition in shrimp GIT, thus maintaining sustainability in the shrimp aquaculture industry.     

The effect of dietary protein level on pond water quality and feed utilization efficiency of pacúPiaractus mesopotamicus (Holmberg, 1887)

The effect of supplemental feeds with different levels of crude protein on pond water quality and food utilization efficiency by pacú (Piaractus mesopotamicus) was evaluated in a semi‐intensive culture system. Fish were stocked at a density of one individual per m², raised for 299 days, and fed isocaloric diets containing soybean meal, blood and bone meal, and yellow maize. Dietary treatments consisted of diets containing 25%, 35%, and 45% crude protein. Ponds receiving the two highest protein levels showed significantly higher values of alkalinity, conductivity and nitrites. High dietary protein diets resulted in no significant improvements in final weight (336.4±77.2, 308.2±92.6 and 368.4±82.2 g, mean±standard deviation, for 25%, 35% and 45% protein levels respectively). Moreover, fish carcass composition was not significantly different among treatments. Instead, significant augmentations in feed conversion ratio (1.36, 1.54 and 1.73 respectively) and reductions in protein retention (35.7%, 33.5% and 29.0% respectively) occurred as protein levels increased. Results indicate that at the lowest protein level, pacú growth rates and carcass composition were similar to the other experimental treatments, with a significant improvement in pond water quality and feed utilization efficiency.     

Elevated α-esterase levels associated with permethrin tolerance in Aedes aegypti (L.) from Baja California, Mexico

A discriminating dose for permethrin for Aedes aegypti from Baja California, Mexico was determined, and was used to select individuals. Mosquitoes were collected from four different municipalities located in the north and south end of the Baja California Peninsula. Individuals were chosen for further study based on their similar response to the insecticide. We exposed 10 groups of 90 Ae. aegypti females to the discriminating dose (172 μg/ml) and after producing 50% mortality, individuals were divided into two categories: killed and survivors. Each of these groups was dissected to separate the head, thorax, and abdomen. Biochemical tests were performed on the head and thorax to determine resistance-related enzyme activities including: α- and β-esterases, glutathione S-transferase, acetyl cholinesterase, insensitive acetyl cholinesterase, and mixed-function oxidases. The results were compared with those for the susceptible New Orleans strain of Ae. aegypti. All the populations studied showed the consistent presence of α-esterases, with elevated levels in permethrin-selected populations. Although β-esterases and GST levels were present in high proportions, they did not reveal a clear pattern in relation to resistance.     

Induction and survival of hemoglobin-less and erythrocyte-less tadpoles and young bullfrogs

Injection of two 25-microgram-per-gram doses of the hemolytic agent phenylhydrazine reduced the hemoglobin level and the erythrocyte count to less than 1 percent of normal tadpole and young blullfrog blood. These anemic animals survive for weeks with little change in overall metabolism. A slow recovery of hemoglobin levels was observed. The implications of this observation for comparative biochemistry are considered.     

Improved assessment of mycelial growth stimulation by low doses of mefenoxam in plant pathogenic <Emphasis Type="Italic">Globisporangium</Emphasis> species

Globisporangium Uzuhashi, Tojo & Kakish. (syn. Pythium Pringsheim) species cause many plant diseases, including Pythium damping-off, leaf and fruit blights, and root rots. Fungicide resistant isolates are selected by repeated use of a single active ingredient on infected crops without rotation. Previous studies demonstrated increased pathogenicity and radial growth in a mefenoxam resistant isolate of Pythium aphanidermatum when exposed to sub-lethal doses of fungicides and ethanol. In those studies, reproducibility of in vitro assays was difficult to achieve due to large variations among trials. This study aimed to examine two protocols for improved reproducibility during the assessment of biphasic dose-responses in mefenoxam-resistant isolates of Globisporangium ultimum and G. irregulare. Two different growth related endpoints, total growth area and total dry mass weight, were assessed. Assays were conducted using ten concentrations of mefenoxam ranging from 0.01 to 1,000 μg/ml. Statistically-significant stimulatory effects were observed in the two Globisporangium species using the two growth related endpoints. Because of its better reproducibility, mycelial growth area is recommended as an endpoint for future studies of chemical hormesis on growth of Globisporangium spp.     

Prevalence and anatomopathological characterization of cutaneous squamous cell carcinomas with regional and distant metastases in dogs and cats: 20 cases (1985–2020)

The aim of this study was to assess the prevalence of regional and distant metastases from cutaneous squamous cell carcinomas (SCCs) in dogs (n = 11) and cats (n = 9) in a retrospective case series over 36 years (1985–2020), as well as to characterize its macroscopic aspects (location and size), degree of differentiation (well, moderately and poorly differentiated [WD, MD and PD, respectively]) and the rate of cell proliferation, by counting the AgNORs. Immunohistochemistry (IHC) was used to identify patterns of tumour migration and invasion (islands, ribbons, cords, small aggregates, individual cells [fusiform and amoeboid]) and to evaluate the intensity of desmoplasia and the amount of myofibroblasts. The prevalence of metastatic SCCs was 4.39% (21/478), being 3.8% in dog (12/309) and 5.3% in cat (9/169). Metastases affected lymph nodes in all dogs and 66% (6/9) of cats, and less frequently distant organs. Primary tumours predominantly affected the abdominal skin in dogs and the nasal planum in cats. Among the 20 cases, 52% were MDs, 34% were WDs, and 14% were PDs. Histological lesions suggestive of exposure to chronic solar radiation were present in 57% (8/14). The main patterns of tumour migration and invasion were islands for WD SCCs and individual cells for PD SCCs. MD SCCs had a mix of patterns. In cats, individual spindle cells were restricted to PDs. A marked desmoplastic reaction was more associated with PD SCCs and often with MDs. This study highlights that the prevalence of SCC metastases in dogs and cats is predominantly regional. The IHC was essential in the identification of individual fusiform keratinocytes, whose presence in surgical margins may represent a greater risk of recurrence. Although the presence of myofibroblasts was observed in all infiltrative and metastatic tumours, further studies evaluating these cells may be important to better understand their role in the tumour microenvironment of cutaneous SCCs with metastasis in dogs and cats.     

Quantification and Characterization of Food Processing Wastes/Residues

A survey was undertaken to determine the wastes/residues generated by the food processing industry in the state of Iowa by sending a questionnaire to food processors with more than ten employees. Food processors were asked to provide a company profile, information about food waste/residue generated, and information about current disposal methods. Samples of wastes/residues were obtained from food processors willing to provide them, and proximate analysis was done. The overall response rate was 82.2 percent or 365 respondents out of 444 questioned. Companies under different category of standard industrial code (SIC) could produce similar wastes/residues that were classified in the same waste/residue streams. Within the meat and seafood category, the majority (87.7 percent) of the wastes/residues were generated by SIC codes 2077 (animal, marine fats and oils) and 2015 (poultry slaughtering, dressing, and processing). Among the waste management practices reported, excluding “other,” land application accounted for 14.0 percent of the waste stream, followed by rendering with 11.4 percent, landfill with 6.7 percent, sewage on-site with 5.0 percent and animal feed production off-site with 4.3 percent. Other waste management practices (combinations of major practices or others not clearly defined) account for 55.1 percent of the allocation of the waste stream.     

Fusion of Boar Sperm with Nanoliposomes Prepared from Synthetic Phospholipids

Liposomes are artificial membrane vesicles that can be used to test and model the functions and interactions of various biological membranes, or as a carrier system to deliver biologically active substances into the cells, or to incorporate lipids into the plasma membrane of target cells to modify membrane structure–function relationships. Sperm plasma membrane undergoes lipid modification during maturation in epididymis and during capacitation in the female reproductive tract to facilitate fertilization. Natural variation in the amounts and composition of lipids in the sperm plasma membrane may also contribute to the species‐specific sperm sensitivities to handling and storage conditions. Boar sperm are notoriously susceptible to membrane damage and are resistant to compositional alteration by artificial liposomes. This study used flow cytometry to demonstrate stable incorporation of nanoliposomes prepared from a complex mixture of various phospholipids (phosphatidylcholine : phosphatidylethanolamine : sphingomyelin : phosphatidylserine : phosphatidylinositol) with high fusion efficiency. Over 90% of sperm rapidly took up fluorescently labelled liposomes and retained the lipids for at least 60 min, in a significant time‐ and concentration‐dependent manner. This unique fusion efficacy could be used to alter sperm plasma membrane composition and hence membrane‐based functional responses.