A histological study of the hph-1 mouse mutant: an animal model of phenylketonuria and infantile hypertrophic pyloric stenosis

AIM: To quantify the chronological sequence of changes in the morphology and immunoreactivity for neurotransmitters in the pylorus of an animal model of infantile hypertrophic pyloric stenosis and phenylketonuria. METHOD: Thirty specimens of pylorus from hph-1 mice and age/sex matched controls (age range: 10-180 days) were examined using conventional histology and immunohistochemistry for a variety of antigens: protein gene product 9.5, a pan neuronal marker; vasoactive intestinal polypeptide; nitric oxide synthase two antigens coalesced to the same inhibitory neurons in humans; substance P, a potent excitatory neurotransmitter; and calcitonin gene related peptide, a neurotransmitter implicated in the somatic afferent innervation of the stomach. The changes in the morphology of the muscle layers were quantified and statistically analysed for each age group (10, 20, 40, 90 and 180 days). RESULTS: Between 10 and 90 days of age, all muscle layers of the hph-1 mice were hypertrophied, for example, 10 days, hph-1 longitudinal muscle mean diameter = 3.4, control = 1.8; hph-1 circular muscle width = 11.5, control = 4.7. The hph-1 mice were significantly smaller during this period (40 days, hph-1 weight = 10 g, control = 25 g). There was no change in the pattern of expression of the antigens examined within the hph-1 mice compared with the controls. CONCLUSION: Hph-1 mice develop a transient smooth muscle hypertrophy of the pylorus attended by gastric distension and failure to gain weight. These changes resolve as the pyloric muscle hypertrophy resolves.     

Transfer of maternal passive immunity to kids in goat herd

The efficiency of tranfer of maternal immunity and its infuence on the kids' health was observed in a herd in which kids (n=20) had whole contact with the dam (n=13). The factors associated with dam, kid and human, which influence the efficiency of passive transfer were observed. It was estimated that the single-born kids reached higher serum gamma-globulin level (mean 23.14 g/dm3) than twin kids (mean 18.2 g/dm3) (p < 0.05). The gamma-globulin level was the highest in single-born kids at 48 h, and in twin kids in 24 h of life. The IgG class antibodies to herd-homologous strains of Pasteurella multocida and Escherichia coli were estimated using ELISA in sera, colostrum and milk whey samples of dams and sera of kids. It was found that maternal antibodies specific to these two facultative pathogens decreased in kids sera rapidly and the self humoral immune response occurred within the period of observation. Two kids delivered by goats with lowest hierarchic position in the herd showed failure of passive transfer, and died at the 10th and 12th weeks of life due to chronic infections induced by both above mentioned bacterial strains.     

Physiological mechanisms of ovarian follicular growth in pigs--a review

Follicular growth after antrum formation is determined by follicle-stimulating hormone (FSH). Only two ways are possible for recruited follicles, continuing development or atresia. In gilts, intensive ovarian follicular growth begins between 60 and 100 days of age, and fluctuations of the ovarian morphological status last about 20 days; however, at that time there are no really large follicles. Final follicular development is under luteinising hormone (LH) control; this is why the attainment of puberty is related to an increase in serum oestradiol to a level that causes a preovulatory surge of this gonadotropin. The pool of follicles at the beginning of the oestrous cycle is about 30-40, most of which are small (< 3 mm) and growing. Then, the pool of follicles increases to about 80 in the mid-luteal phase but about 50 of them are small and 30 are medium sized (3-6.9 mm). Some of these follicles are in the growing phase, but some are atretic. Between days 7 and 15 of the oestrous cycle the percentage of atretic follicles fluctuates between 12 and 73%. At that time there are no large (> 7 mm) follicles because of the suppressing effect of progesterone. The number of small follicles declines after luteolysis. From the pool of medium follicles, large follicles are selected under the influence of LH, but about 70% of the medium-sized follicles become atretic. Because of the long-lasting selection process there is a significant heterogeneity in the diameter of large follicles in oestrus. However, the number of follicles correlates with the number of corpora lutea after ovulation. Individual follicular development and the relationship between follicles are still poorly known. The use of ultrasonography may give a closer insight into these phenomena.     

Occurrence of the structural enterocin A, P, B, L50B genes in enterococci of different origin

Enterococci are well-known producers of antimicrobial peptides--bacteriocins (enterocins) and the number of characterized enterocins has been significantly increased. Recently, enterocins are of great interest for their potential as biopreservatives in food or feed while research on enterocins as alternative antimicrobials in humans and animals is only at the beginning. The present study provides a survey about the occurrence of enterocin structural genes A, P, B, L50B in a target of 427 strains of Enterococcus faecium (368) and Enterococcus faecalis (59) species from different sources (animal isolates, food and feed) performed by PCR method. Based on our results, 234 strains possessed one or more enterocin structural gene(s). The genes of enterocin P and enterocin A were the most frequently detected structural genes among the PCR positive strains (170 and 155 strains, respectively). Different frequency of the enterocin genes occurrence was detected in strains according to their origin; the strains from horses and silage showed the highest frequency of enterocin genes presence. All possible combinations of the tested genes occurred at least twice except the combination of the gene of enterocin B and L50B which possessed neither strain. The gene of enterocin A was exclusively detected among E. faecium strains, while the gene of enterocin P, B, L50B were detected in strains of both species E. faecium and E. faecalis. In conclusion, a high-frequency and variability of enterocin structural genes exists among enterococci of different origin what offers a big possibility to find effective bacteriocin-producing strains for their application in veterinary medicine.     

Goblet cells and mucus layer in the gut of young pigs: Response to dietary contents of threonine and non‐essential amino acids

Mucins secreted by goblet cells (GC) are the major components of mucus layer coating and protecting gut epithelium. The study aimed at determining the effect of non‐essential amino acids (NEAA) and threonine (Thr) levels on GC number and mucus layer thickness measured in different parts of the gut. A two‐factorial experiment was conducted on 72 pigs (initial BW 12.5 kg) using as factors: Thr level (5.1, 5.7, 6.3 and 6.9 g standardized ileal digestible [SID] Thr/kg) and wheat gluten (WG) level used as a source of NEAA (20.4, 40.4 and 60.4 g WG protein in WG20, WG40 and WG60 diets respectively). All diets covered the requirement for essential AA, except for Thr, and they were fed to six pigs for 20 days. Thr level affected only the count of GC containing acidic mucins in the proximal colon, which was higher in pigs fed a diet with 5.1 g SID Thr/kg in comparison with diet containing 6.3 g SID Thr/kg. In the villi, WG40 and/or WG60 increased the GC number containing acidic mucins and lowered that with neutral mucins in the middle jejunum. In the crypts, higher WG levels decreased the GC number with acidic mucins in the duodenum, ileum and proximal colon and increased that with neutral and mixed mucins in the duodenum and ileum. The mucus layer was thicker in the proximal and middle jejunum in WG20 pigs, whereas in the proximal colon it was thicker in WG60 pigs. The lowest GC count was found in the middle jejunum and the highest in the duodenum and proximal colon. The mucus layer was the thinnest in the proximal and middle jejunum and the thickest in the ileum. The results did not show the negative effect of a moderate Thr deficiency and the positive effect of non‐essential AA supplementation on GC functions.     

Proteome changes in renal cortex and medulla induced by dietary supplementation with inulin‐type fructans in growing pigs

The aim of the study was to evaluate the influence of dietary supplementation with inulin extract from chicory root and dried chicory root on the protein profile of the renal cortex and medulla of growing pigs. The experiment was carried out on renal cortex and medulla tissue collected from 24 50‐day‐old PIC x Penarlan P76 crossbred piglets (males). Animals were divided into three dietary groups (n = 8) and fed with a control diet, diet supplemented with 2% inulin extract from chicory root and a diet supplemented with 4% dried chicory root. Kidney samples were collected after 40 days of feeding, and renal cortex and medulla proteins were separated by two‐dimensional electrophoresis. Protein identification was performed using MALDI‐TOF mass spectrometry. The diet supplemented with 2% chicory inulin induced significant expression changes of 20 and 26 protein spots in the renal cortex and medulla respectively. Supplementation with 4% dried chicory root triggered changes in the expression of 44 and 24 proteins in the renal cortex and medulla respectively. Both forms of chicory inulin‐type fructans effectively affected the expression of proteins involved in energy metabolism, heat shock proteins and other chaperones, cytoskeletal and cytoskeleton‐related proteins, as well as other proteins. Additionally, changes in transferrin abundance in both experimental groups suggested the significance of chicory fructan supplementation for iron absorption and bioavailability. In conclusion, 2% inulin extract from chicory root and 4% dried chicory root exerted a similar effect on changes in renal protein expression; however, more pronounced alterations were induced by dried chicory root. Nevertheless, further studies are needed for better understanding the mechanism underlying the effect of chicory inulin‐type fructans and their fermentation end products on the kidneys of growing pigs.     

Alterations in the rainbow trout (Oncorhynchus mykiss) eggs exposed to ionizing radiation during induced androgenesis

Ionizing radiation (IR) is applied to inactivate nuclear genome in the salmonid eggs to induce androgenetic development. However, it has been considered that doses of IR used to damage maternal chromosomes may also affect morphology of the eggs and decrease their developmental potential. Thus, the main goal of the present research was to assess alterations in the rainbow trout (Oncorhynchus mykiss) eggs caused by the high dose of IR administered during androgenesis. In the present research, rainbow trout eggs were irradiated with 350 Gy of X‐rays, inseminated and exposed to the high hydrostatic pressure (HHP) shock to develop as androgenetic doubled haploids (DHs). The distribution of lipid droplets in the irradiated and non‐irradiated rainbow trout eggs, survival rates and morphology of larvae from androgenetic and control groups were compared. It has been observed that non‐irradiated and irradiated eggs exhibited altered distribution of lipid droplets. Most of the eggs before IR treatment displayed rather equal distribution of the oil droplets. In turn, majority of eggs studied after irradiation had coalesced lipid droplets, a pattern found in eggs with reduced quality. Incidences of abnormally developed larvae were more frequently observed among fish that hatched from the irradiated eggs. Observed changes suggest X‐rays applied for the genetic inactivation of rainbow trout eggs may lead to decrease of their developmental competence.     

Is the consumption of snail meat actually healthy? An analysis of the osteotropic influence of snail meat as a sole source of protein in growing rats

The study was aimed at determining the osteotropic effects of diets containing snail meat as a sole protein source. In our experiment, we tested three different diets incorporating snail meat originating from Helix pomatia (HP), Cornu.aspesa maxima (CAM) and Cornu.aspersum aspersum (CAA) and compared these to a control diet (CON) in which casein was the source of protein. In all diets, the protein content amounted to 10%, as calculated on a dry weight basis. In the study, forty male Wistar rats with an initial body mass of 50 ± 2 g were randomly placed within the control and three experimental groups. After 28 days of experimental feeding, the rats were sacrificed, and the body mass, total skeletal density and body composition were recorded. Moreover, blood serum (osteocalcin, CTX) and isolated tibia (pQCT, DXA, 3D micro‐CT, 3‐point bending test) were stored for further analysis. The results reveal that a diet incorporating snail meat significantly decreased BMC (bone mineral content), as well as area of total skeleton and isolated tibia, and was without influence on BMD (bone mineral density). Furthermore, the 3D micro‐CT analysis of trabecular compartment documented a reduced Tb.Th (trabecular thickness), as well as Tb.N (trabecular number), and an increased Tb.Sp (trabecular separation). Beyond the aforementioned, the snail‐based diets had an influence upon the architectonical properties of the tibia—decreasing its resistance to mechanical loading. Finally, snail meat, when used as an alone source of protein, negatively influenced the metabolism of the bone tissue in growing animals—making bone smaller and weaker.     

Modulation of ruminal and intestinal fermentation by medicinal plants and zinc from different sources

Two experiments were conducted on sheep to determine the effect of dietary supplementation with zinc and a medicinal plant mixture on haematological parameters and microbial activity in the rumen and large intestine. In Experiment 1, 24 male lambs were randomly divided into four groups: One group was fed an unsupplemented basal diet (control), and three groups were fed a diet supplemented with 70 mg Zn/kg diet in the form of Zn sulphate (ZnSO₄), a Zn‐chelate of glycine hydrate (Zn‐Gly) or a Zn‐proteinate (Zn‐Pro), for five months. The ruminal content was collected separately from each lamb, and batch cultures of ruminal fluid were incubated in vitro with mixture of medicinal plants (Mix) with different roughage:concentrate ratios (800:200 and 400:600, w/w). Bioactive compounds in Mix were quantified by UPLC/MS/MS. In Experiment 2, four sheep were fed a diet consisting of meadow hay and barley grain (400:600, w/w), with Zn‐Gly (70 mg Zn/kg diet), Mix (10% replacement of meadow hay) or Zn‐Gly and Mix (Zn‐Gly‐Mix) as supplements in a Latin square design. Mix decreased total gas (p < 0.001) and methane (p < 0.01) production in vitro. In Experiment 1, caecal isobutyrate and isovalerate concentrations varied among the dietary treatments (p < 0.01). The isovalerate concentration of the zinc‐supplemented groups in the distal colon was higher (p < 0.001) compared with the control. In Experiment 2, the molar proportion of isobutyrate was the highest in the faeces of the sheep fed the diet with Zn‐Gly‐Mix (p < 0.01). The plasma zinc concentration was higher in the groups fed a diet supplemented with zinc (p < 0.001). The haematological profile and antioxidant status did not differ between the dietary groups (p > 0.05). The diets containing medicinal plants and organic zinc thus helped to modulate the characteristics of fermentation in ruminants.