Dry preservation of Toxocara vitulorum by plastination technique

Background and Aim: The most widely adopted technique to preserve the gross specimen of the parasite is immersions and storage in liquid preservatives. The present study aimed to describe the dry method of the preservation of Toxocara vitulorum using plastination technique. Materials and Methods: Acetone dehydrated parasites were incubated at –20°C for 1 month in five different plastination solutions, prepared by mixing melamine and turpentine oil with clove oil (MTCl)/chloroform (MTC)/isopropanol (MTI)/benzene (MTB)/xylene (MTX) in 1:1:1 ratio to infiltrate the polymer. Technical personnel was asked to assign weekly score for dryness, stickiness, shrinkage, glossiness, flexibility, and odor of the prepared model on a 5-point scale. Results: Overall, the plastinated parasites were dry, non-sticky, glossy, odorless, chemical-free, harmless, to some extent flexible, with detectable morphological structure including natural form but lost their natural color, and cuticle became translucent. A varying level of shrinkage was noted in all types of plastinated model, but it was least in MTCl model. One month post-plastination, the mean evaluation score for glossiness was maximum in the parasite plastinated in MTCl solution (4.50±0.17), followed by MTC (3.72±0.32), MTX (3.56±0.38), MTB (2.83±0.37), and MTI (2.31±0.33). Likewise, for flexibility, the score was maximum in the parasite plastinated in MTCl solution (4.36±0.16), followed by MTB (3.11±0.14), MTC (2.94±0.41), MTX (2.75±0.41), and MTI (1.97±0.28). The degree of dryness, stickiness, and odor of the prepared model varies non-significantly (p>0.05) with the polymer mixtures. Maximum shrinkage percentage in terms of length and width was 4.24% and 50%, respectively, in the parasites plastinated in MTB solution. Shrinkage percentage was minimal (1.81% in length and 25% in width) in the MTCl plastinated parasites. Shrinkage percentage in terms of dimension was statistically non-significant among the different polymer solutions. Plastinated models withstand the process of microbial decomposition. There were 5 and 11 odd points in favor of plastination and formalin preservation technique, respectively. Conclusion: The prepared T. vitulorum model in MTCl can be used as an adjunct to the parasite preserve in 10% formalin solution. The plastination technique can be used as an alternative method of liquid preservation.

Hematological parameters in largemouth bass (Micropterus salmoides) with formalin-preservation: comparison between wild tournament-caught and captive-raised fish

Background Largemouth bass (Micropterus salmoides) are an economically important freshwater fish species that have been investigated for both the short and long-term effects of stress, secondary to angling. Limited data has been published on the hematological parameters of this species and blood sample stability is a notable limitation of hematologic field studies. A relatively novel technique using 10% neutral buffered formalin preserves heparinized whole blood and maintains blood cell stability beyond one month in striped bass. The objective of this study was to evaluate the differences in hematological parameters between tournament-caught and captive-raised largemouth bass using whole blood preservation with neutral buffered formalin. Methods Two populations of largemouth bass (n = 26 wild; n = 29 captive) underwent coccygeal venipuncture to collect heparinized whole blood for packed cell volume, total solids, and manual differential. Formalin preservation of heparinized whole blood facilitated manual hemocytometer analysis. Results were compared between the populations (tournament-caught, and captive-raised) with Wilcoxon rank sum test, a Hotelling’s T2 test, and Bonferroni simultaneous 95% confidence intervals to determine significance. Results The mean packed cell volume (44.9 ± 5.4%) and total solids (7.2 ± 1.1 g/dL) were significantly higher, while the total leukocyte count (7.08 ± 1.86 × 103/µL) was significantly lower in the wild tournament-caught population of largemouth bass, as compared to the captive-raised counterparts (PCV 34.4 ± 7.2%; TS 5.2 ± 1.0 g/dL; WBC 16.43 ± 8.37 × 103/µL). The wild population demonstrated a significantly distinct leukogram characterized by a neutropenia (24.1 ± 12.7%), lymphocytosis (67.7 ± 13.0%), and monocytopenia (8.3 ± 2.9%), while the erythrocyte and thrombocyte counts were not significantly different between populations. Discussion Numerous factors have been demonstrated to influence hematologic parameters in fish including age, size, sex, temperature, environmental oxygen level, population density, and infection. The wild population endured stress during angling capture, live-well hypoxia, transport, and extended air exposures at weigh in, which may have caused a stress leukopenia as well as osmoregulatory dysfunction and subsequent hemoconcentration. Further evaluation of seasonal impact as well as increased sample size is warranted to enhance our understanding of largemouth bass hematology. Conclusion This study concluded that wild largemouth bass captured via tournament angling have higher packed cell volume and total solids, and lower total leukocyte counts, compared to captive-reared individuals. Through the completion of this study, we demonstrated the successful use of 10% neutral buffered formalin to preserve heparinized whole blood for precise hemocytometer cell counts in a new teleost species, the largemouth bass.

Eksplorasi Rafinosa Biji Kapas Sebagai Pengganti Formalin Dalam Pengawetan Ikan [Raffinose Exploration Cotton Seeds Lieu Of Formalin In Preserving Fish]

Preservation is a human effort to enhance durability and shelf life of the fish that the quality of fish remains in good condition. Irregularities in the use of formalin preservation would endanger the health of consumers. Cottonseed is containing compounds of Raffinose, proteins, fats that are cryoprotectants to cells and antimicrobe, that use of cottonseed to suppress use of formalin as a preservative of fish in the community. The method used by dissolving cottonseeds in organic solvents, where in the organic solvent effective to extract the active ingredients in cottonseed. The results showed use of cotton seed extract is 100% equal to 100% use of formalin in maintaining the quality of the fish. This is evidenced by organoleptic tests, pH and bacterial tests. Expected with these results, cottonseed can be a substitute for use of formalin.