Therapeutic Intervention with Dietary Chitosan Nanoparticles Alleviates Fish Pathological and Molecular Systemic Inflammatory Responses against Infections

Marine bio-sourced chitosan nanoparticles (CSNP) are antimicrobial and immunomodulatory agents beneficial for fish medicine. Herein, dietary CSNP was investigated for the amelioration of the systemic inflammatory responses of an induced fish model. One hundred and forty-four rainbow trout were assigned to one pathogen-free and non-supplemented group (negative control), and three challenged groups: non-supplemented (positive control), CSNP-preventive, and CSNP-therapeutic. After a feeding experiment extended for 21 days, the organosomatic indices (OSI) and molecular aspects were assessed. After a challenge experiment extended for further 28 days, CSNP-therapeutic intervention was assessed on fish survival and systemic inflammatory responses on pathology, histo-morphology, and molecular aspects. With CSNP administration, OSI nonsignificantly decreased and the relative expression of targeted inflammatory-mediator genes was significantly increased. The CSNP-therapeutic fish showed an RPS of 80% as compared to the positive control group, and CSNP-therapeutic administration retained the highest gene expression augmentation up to 28 days after the challenge. Notably, the splenic reticulin fibers framework of the CSNP-therapeutic group retained the highest integrity among the groups during the infection. After recovery, reticulin fibers density in the CSNP-therapeutic samples was significantly higher than in the negative control group, which indicates high innate immunity. Thus, CSNP showed promising biotherapeutic features enhancing fish resistance against infections.     

Abortion rate and mortality among exotic pregnant heifers imported to the Gezira in the central region of the Sudan

Despite the intensive management and husbandry precautions, exotic pregnant heifers imported to the Gezira, Sudan, suffered excessive stress. Four of five heifers aborted due to stress. Death was due to pasteurellosis, babesiosis and heart water.     

Yeasts and algae isolated from cows with mastitis in the south-eastern part of Poland

The aim of the study was to evaluate the occurrence of mycotic and protohecal mastitis in herds in south-eastern part of Poland. A total of 3091 milk samples from udder quarters with clinical and subclinical mastitis from 29 dairy herds was investigated in this survey. Milk samples were plated as soon as possible on blood agar (BA), Mac Conkey agar, aesculin-tallium acetate-crystal violet agar, and Sabouraud agar. A hundred and thirty one yeast (4.23%) and eleven Protoheca zopfii (0.35%) strains were isolated from cows with clinical and subclinical mastitis. All the isolated fungi were the yeast classified into 4 genera (Candida, Trichosporon, Saccharomyces and Rhodotorula). The most frequently isolated yeasts were Candida sp., C. kefyr, C. humicola, C. rugosa and C. inconspicua. Both fungi and algae were isolated first of all during a confinement-housing season.     

Immune dynamics following infection of avian macrophages and epithelial cells with typhoidal and non-typhoidal Salmonella enterica serovars; bacterial invasion and persistence, nitric oxide and oxygen production, differential host gene expression, NF-κB signalling and cell cytotoxicity

Poultry-derived food is a common source of infection of human with the non-host-adapted salmonellae while fowl typhoid and pullorum disease are serious diseases in poultry. Development of novel immune-based control strategies against Salmonella infection necessitates a better understanding of the host-pathogen interactions at the cellular level. Intestinal epithelial cells are the first line of defence against enteric infections and the role of macrophages is crucial in Salmonella infection and pathogenesis. While gene expression following Salmonella infection has been investigated, a comparison between different serovars has not been, as yet, extensively studied in poultry. In this study, chicken macrophage-like cells (HD11) and chick kidney epithelial cells (CKC) were used to study and compare the immune responses and mechanisms that develop after infection with different Salmonella serotypes. Salmonella serovars Typhimurium, Enteritidis, Hadar and Infantis showed a greater level of invasion and/or uptake characters when compared with S. Pullorum or S. Gallinarum. Nitrate and reactive oxygen species were greater in Salmonella-infected HD11 cells with the expression of iNOS and nuclear factor-κB by chicken macrophages infected with both systemic and broad host range serovars. HD11 cells revealed higher mRNA gene expression for CXCLi2, IL-6 and iNOS genes in response to S. Enteritidis infection when compared to S. Pullorum-infected cells. S. Typhimurium- and S. Hadar-infected HD11 showed higher gene expression for CXCLi2 versus S. Pullorum-infected cells. Higher mRNA gene expression levels of pro-inflammatory cytokine IL-6, chemokines CXCLi1 and CXCLi2 and iNOS genes were detected in S. Typhimurium- and S. Enteritidis-infected CKC followed by S. Hadar and S. Infantis while no significant changes were observed in S. Pullorum or S. Gallinarum-infected CKC.     

Genetic Structure of Severe Combined Immunodeficiency Carrier Horses in Morocco Inferred by Microsatellite Data

A total of 17 microsatellite deoxyribonucleic acid loci used routinely for horse parentage control were used to evaluate genetic diversity among normal Arabian horses and severe combined immunodeficiency (SCID) carrier Arabian horses (ArS) and normal Arab-Barb horses and SCID carrier Arab-Barb horses (ArbeS). On the basis of the genotype of 186 horses, mean allelic diversity was estimated as 6.82, 5.53, and 6.7059 in normal Arabian horses, ArS, and for both groups of Arab-Barb horses, respectively. Five specific alleles were observed in ArS and ArbeS, with one common with ArS at HMS6, whereas five alleles common between ArS and ArbeS had a high frequency. Expected and observed heterozygosity showed great heterogeneity in the population studied and were similar or higher when compared with other studies on Arabian horses. Coefficient of gene differentiation G_st of Nei associated with Nei’s genetic distance and multivariate correspondence analysis indicated a possible differentiation between the studied populations when analyzed separately according to breed. Probability of assignment of a horse to a specific group was assessed using a full and partial Bayesian approach. In all, 80.6% of Arab horses and 78.2% of Arab-Barb horses were assigned properly with a partial Bayesian test, which provided better results than the full one. These findings will be useful for identification of SCID carrier horses by using the microsatellite deoxyribonucleic acid loci used routinely for horse parentage control in our laboratory.     

The detection of the meq gene in chicken infected with Marek's disease virus serotype 1

In the genome of strains of very virulent Marek's disease virus serotype 1(vvMDV1), such as Md5 and RB1B, the meq open reading frame (ORF) encoding a 339-amino-acid bZIP protein, is present, while a slightly longer meq ORF, termed as L-meq, in which a 180-bp sequence is inserted into the meq ORF is found in other strains of MDV1, such as CV1988/R6 and attenuated JM. When chickens were infected with vvMDV1 strains and the meq gene was amplified by nested polymerase chain reaction (PCR), the meq gene was detected throughout the experimental period for 7 weeks post inoculation (pi). However, the L-meq gene was also detected at 3 to 5 weeks and 3 to 4 weeks pi. in Md5-infected and RB1B-infected chickens, respectively. In the case of chickens infected with an attenuated MDV1, the JM strain, the L-meq gene was detected at 2 to 7 weeks pi., and the meq gene was also detected at 2 to 6 weeks pi. Both L-meq and meq genes were detected in chickens infected with an attenuated nononcogenic vaccine strain of MDV1 (CVI988/R6), throughout the experimental period. Though quantitative PCR was not performed, a larger amount of the PCR products corresponding to the L-meq than the meq gene was amplified from chickens infected with JM or CVI988/R6. These results suggest that a dynamic population shift between the MDV subpopulations displaying meq and L-meq genes occurs in chickens during the course of MDV infection. Since the MDV subpopulation that displays the L-meq gene only displays it during the latent phase, the L-meq and its gene product, if any, might contribute to the maintenance of the MDV latency.     

Abomasal cannulation in the milk-fed calf using a 7 mm polyurethane tube

The main objective of this study was to develop a simple and effective surgical technique for abomasal cannulation in neonatal calves. General anaesthesia was induced in 12, 3-day-old male dairy calves and a polyurethane cannula surgically implanted in the abomasal body (n = 12) and pyloric antrum (n = 6) through a right paracostal incision. Fifteen cannulae remained in situ from day 3 to 34 of life (mean: 29 days), and three cannulae were extruded 13-14 days after placement. Calves were clinically healthy and gained weight during the study. Cannulae were well tolerated by the calves and abomasal contents did not leak from the cannula sites. Necropsy examination revealed firm adhesions between the abomasum and parietal peritoneum at the cannula sites with no evidence of leakage or peritonitis. We conclude that surgical placement of polyurethane tubes designed for percutaneous endoscopic gastrostomy provided a useful method for cannulation of the abomasum of neonatal calves. The cannulation technique can be used for experimental studies, as well as for nutritional and fluid support of sick calves that cannot be managed by oral treatment.     

Sonographic characteristics of goat testis on water bath based ultrasonography

Assessing the health of the testes in domestic animals is an important aspect of the breeding soundness examination and selection. The aim of the present study was to develop a simple method for scanning and to establish ultrasonographically the gross anatomic structures of the goat testes. Six adult male goats were examined to study the sonographic appearance of normal testes and epididymides using a water bath based ultrasound scanning technique. The ultrasonographic examinations were done using a 5–9 MHz/60 mm (7.5 MHz) linear-array transducer and a B-mode scanner. The ultrasonographic examination was performed in goats after standardizing the procedure on six testes collected from slaughter house. Results showed that in live goats when the probe was placed directly over the scrotum it gave distorted and unclear image. In water bath method the entire scrotum was dipped into a container filled with water and linear probe was used to observe the sonographic features of the testis. Each testis was viewed vertically, resulting in longitudinal image which was frozen, measured and printed through a thermal printer. The results of the ultrasonogram revealed that the testicular parenchyma was homogenous and moderately echogenic throughout. The diameter (mean±se) of the right and left testes was 4.47±0.14 and 4.42±0.07 cm respectively and no significant difference was observed between the testes. The mediastinum testis was a 1.50±0.22 cm wide linear structure of greater echogenicity than the testicular parenchyma when viewed in the transverse plane and nearly circular echogenic “spot” in the midline of the testis when viewed horizontally. The head and tail of the epididymides were easily identified on all the testes, but the epididymal body and ductus deferens were difficult to identify consistently. The tail of the epididymis was easily identified on the distal end of the testis with sonolucent tubules and appeared sonographically as a ‘peaked cap’ upon the testicular parenchyma. The diameter (mean±se) of the tail of right and left epididymis was 2.11±0.18 and 1.92±0.06 cm and no significant difference was observed between epididymides. The vascular pampiniform plexus (1.42±0.18 cm) was easily identified on the proximal end of the testes. The tunics of the testes appeared as a bright echogenic line. Inter-testicular septum appeared between testes as a hyperechoic line. It is concluded that ultrasonography permits a noninvasive evaluation of the internal structure of the scrotum and testes and water bath based sonographic examination may prove to be a valuable simple diagnostic methodology for evaluating physiopathologic conditions of goat testes and can be employed as a routine investigative method during breeding soundness and clinical examination.     

Effects of leptin and tumor necrosis factor-alpha on degranulation and superoxide production of polymorphonuclear neutrophils from Holstein cows

Leptin, a pleiotropic hormone regulating food intake and energy expenditure, has been shown to directly modulate human polymorphonuclear neutrophil (PMN) functions or indirectly through the action of tumor necrosis factor-alpha (TNF-alpha). Bovine PMN have considerable different characteristics from human PMN. For example, it does not respond to N-formyl-Methionyl-Leucyl-phenylalanine, a well known human PMN activator. In the present study, we tested the effects of leptin and TNF-alpha on superoxide production and degranulation of bovine peripheral PMN, in which both long isoform of leptin receptor (Ob-Rb) and TNF receptor 1 were expressed. Human leptin, human TNF-alpha, phorbol myristate acetate (PMA) and opsonized zymosan particles (OZP) did not stimulate degranulation responses, while zymosan-activated serum (ZAS) did. Neither leptin nor TNF-alpha enhanced the ZAS-induced degranulation responses. TNF-alpha, PMA, OZP and ZAS increased superoxide production in different magnitudes, whereas leptin did not. TNF-alpha, but not leptin, enhanced OZP- and ZAS-induced superoxide production, possibly, in part due to facilitating translocation of p47(phox), a component of NADPH oxidase. These results indicate that, unlike in human PMN, leptin does not have any direct effect on degranulation and superoxide production in bovine PMN, although TNF-alpha influences superoxide production.