Experimental Infections of Bluegill with the Trematode Ribeiroia ondatrae (Digenea: Cathaemasiidae): Histopathology and Hematological Response

Infections by the digenetic trematode, Ribeiroia ondatrae, cause severe limb malformations in many North American amphibians. Ribeiroia ondatrae also infects fishes as second intermediate hosts, but less is known about the pathology and immune responses initiated in infected fish, even though reports of infected fish date back to early 1900s. To this end, we experimentally exposed juvenile Bluegills Lepomis macrochirus to three doses of R. ondatrae cercariae and monitored the pathology, parasite infection success, and humoral responses over 648 h. All exposed fish became infected with metacercariae, and the average infection load increased with exposure dose. Histologically, infection was associated with acute hemorrhages in the lateral line and local dermis at 36 h, followed by progressive granulomatous inflammation that led to the destruction of encysted metacercariae. Correspondingly, over the course of 648 h we observed an 85% decline in average infection load among hosts, reflecting the host's clearance of the parasite. Infection was not associated with changes in fish growth or survival, but did correlate with leukocytosis and neutrophilia in circulating host blood. Understanding the physiological responses of R. ondatrae in Bluegill will help to clarify the ecological effects of this parasite and provide a foundation for subsequent comparisons into its effects on behavior, individual health, and population dynamics of Bluegill.

Received March 4, 2015; accepted August 9, 2015     

An in vitro method to determine phosphorus digestibility of rainbow trout Oncorhynchus mykiss (Walbaum) feed ingredients

An in vitro method was developed to assess the digestibility of phosphorus in 12 plant and animal feed ingredients for rainbow trout Oncorhynchus mykiss (Walbaum). The method simulates the gastrointestinal tract of the rainbow trout with regard to pH and gastrointestinal enzymes. Phosphorus solubility was measured after acid digestion (pH 3) with and without gastric enzymes, after alkaline digestion (pH 9) with and without intestinal enzymes, and after a two-step process involving acid and alkaline digestion. Commercially available digestive enzymes from mammals were compared with digestive enzymes from rainbow trout. Correlating in vitro digestibility with in vivo digestibility showed that acid digestion with both commercial enzymes ( r ²=0.98, P  < 0.05) and trout enzymes ( r ²=0.94, P  < 0.05) predicted the in vivo digestibility of animal feed ingredients. Alkaline digestion with both enzyme systems (commercial r ²=0.79; trout r ²=0.74, P  < 0.05) or without ( r ²=0.82, P  < 0.05) enzymes predicted the in vivo digestibility of ingredients from animal byproducts but not those from plant products. The in vitro digestibility with two enzyme steps (acid and alkaline) predicted in vivo digestibility of plant and animal ingredients ( r ²=0.79 for commercial enzymes and r ²=0.74 for trout enzymes) better than did one-step acid or alkaline digestion.     

Effect of Breeding Activity on the Microflora of the External Genitalia and in the Semen of Stallions,and the Relationship Between Micro‐organisms on the Skin and on the External Genitalia

A possible role of breeding activities in the composition of the microbial population in stallions' external genitalia (EG) and the relationship between micro‐organisms colonizing the skin of the abdomen and the ones colonizing the EG have not been studied. In experiment 1, EG microbiological samples were collected from 41 stallions used for both natural cover and semen collection (BST) and from 18 non‐breeding stallions (NBST). A higher (p < 0.05) frequency of isolation of potentially pathogenic species was found for BST. Age did not influence number of micro‐organism species isolated both in BST and NBST. In experiment 2, the microbial content of the EG and semen was compared in 23 BST. Most micro‐organisms isolated from the EG were present in semen, albeit with a numerically lower prevalence. In 7 stallions, six microbial species isolated from semen were absent from the EG cultures, suggesting contamination by the operator. In experiment 3, a numerically higher number of micro‐organism species was isolated from the EG of 31 stallions, than from their skin of the ventral abdomen in contact with the penis or from the skin of the thorax. With the sole exception of Escherichia coli, potentially pathogenic bacteria were only isolated from the EG but not from the skin. Results suggest that breeding activity increased the number of species colonizing the EG; most species isolated from the EG were also found in semen even if with a lower frequency, and additional semen contamination seemed to occur during its manipulation. Many micro‐organism species of the skin were also isolated from the penis, but independently of being or not in contact with the penis, skin did not seem to provide an adequate environment for the growth of potentially pathogenic bacteria that were isolated from EG, with the sole exception for E. coli.     

The nutritional properties of co-dried fish silage in rainbow trout (Salmo gairdneri) dry diets

Feed ingredients containing fish silage and liquefied fish made from ground, whole Pacific whiting and co-dried in a vacuum dryer with mixtures of soybean meal and feather meal to facilitate drying were prepared. An additional batch of fish silage was co-dried with the other dry ingredients in the diet formulation that was used, Abernathy diet S8-1. Fish meal, made by vacuum drying Pacific whiting, was used in the control diet. Co-dried fish meal was made by co-drying Pacific whiting with a soybean meal-feather meal mixture. Fish meal was entirely replaced by the co-dried products in the experimental diets, which were fed to rainbow trout (Salmo gairdneri) for 32 weeks. The best growth and food conversion values were obtained by feeding the fish meal control diet or the diet in which the fish meal was replaced with co-dried liquefied fish. No significant differences in final weights were found between trout fed diets containing co-dried fish meal or co-dried fish silage (fish products were 25% of the diet), but these fish were significantly smaller than fish fed the fish meal control or the co-dried liquefied fish diets. Reducing the fish silage to 12.5% or increasing it to 50% further reduced weight gains in the trout. Food conversion values, protein efficiency ratios, and net protein utilization values generally followed the same trends between diets as did the final weight values. Apparent digestibility coefficients for the co-dried products were lower than for the fish meal, possibly because they contained soybean meal-feather meal mixtures. Organoleptic properties of the fish were not affected by diet.     

大豆粕在鲑鳟鱼类饲料中的利用

~~大豆粒在鲑鳟鱼类饲料中的利用@Ronald W.Hardy     

Dietary microbial phytase supplementation and the utilization of phosphorus, trace minerals and protein by rainbow trout [Oncorhynchus mykiss (Walbaum)] fed soybean meal-based diets

Effects of thermal and enzymatic treatments of soybean meal on apparent absorption of total phosphorus, phytate phosphorus, nitrogen (protein), ash, calcium, magnesium, copper, iron, manganese, strontium and zinc were examined using rainbow trout, Oncorhynchus mykiss (Walbaum), as the test species. Absorption of the test nutrients was estimated using yttrium as an inert non-absorbable indicator. Thermal treatments (microwaving, dry roasting, steam heating, cooking) had no measurable effect on the apparent absorption of phosphorus and other minerals. Phytase supplementation increased the apparent absorption of phosphorus, nitrogen (protein), ash, calcium, magnesium, copper, iron, strontium and zinc in low-ash diets containing soybean meal, but had little effect in high-ash diets containing both soybean and fish meal. In low-ash diets, the apparent absorption of phosphorus increased in accord with the level of phytase added to the diet, from 27% (no phytase added) up to 90% (phytase added, 4000 units kg⁻¹ diet) or 93% (predigested with phytase, 200 units kg⁻¹ soybean meal). In high-ash diets, dietary acidification with citric acid decreased the effect of phytase, whereas in low-ash diets, acidification markedly increased the effect of the enzyme. Excretion of phosphorus in the faeces of fish fed a low-ash diet containing phytase-treated soybean meal was 0.32 g per kg diet consumed, a 95%−98% reduction compared with phosphorus excretion by fish consuming commercial trout feeds.     

Apparent digestibility coefficients of nutrients in brewer's and rendered animal by-products for rainbow trout (Oncorhynchus mykiss (Walbaum))

Two experiments were conducted to measure the apparent digestibility coefficients (ADCs) of nutrients, dry matter, and energy in by‐products of the brewing industry and in selected animal protein ingredients for rainbow trout. In experiment 1, 500 rainbow trout (average body weight 170.8±5.5 g) were stocked in ten 140‐L digestibility tanks with 50 fish per tank and two tanks per diet. Yttrium oxide was used as an inert marker in the diets. The high fibre content of brewer's dried grains (BDG) affected the ADCs of dry matter but not of protein or amino acids. Brewer's dried yeast had a higher protein content than BDG, but ADC values for protein and amino acids were significantly lower. The ADCs of phosphorus were similar among brewer's dried products. The BDG high‐protein, fraction had marginally higher ADC values for dry matter, protein and amino acids than regular BDG. Conditions in experiment 2 were similar to those in experiment 1. ADC values for spray‐dried porcine plasma were over 98% for dry matter, crude protein, crude fat, and gross energy. ADC values for spent hen meal were higher than those of poultry by‐product meal or feather meal, with the exception of gross energy. However, spent hen meal was unpalatable. The ADCs in these ingredients were variable, and this variability must be taken into account when these ingredients are formulated into feeds for fish.     

Development and evaluation of a rapid absorbed enzyme immunoassay test for the diagnosis of Johne''s disease in cattle

An absorbed enzyme immunoassay (EIA) test for Johne's disease in cattle was developed in which absorption of cross-reacting antibodies occurred as a rapid reaction in solution rather than overnight with whole organisms and a subsequent centrifugation step. Total test time was reduced to less than 2 h with a minimum of manipulations. The test was evaluated in cattle herds from Johne's disease-endemic and Johne's disease-free regions of Australia. Specificity was 99.8%. Calculations of sensitivity were affected by the history of the herd under test. However, the EIA detected in excess of 80% of animals before onset of clinical disease and 65% of faecal shedders were EIA positive on, or before, first detection of Mycobacterium paratuberculosis in their faeces. The test should aid epidemiological studies and be a useful tool in the management and control of Johne's disease.