Effect of the supplementation with a blend containing short and medium chain fatty acid monoglycerides in milk replacer on rumen papillae development in weaning calves.

Feeding of neonates with artificial milk formulas is a popular trend toward early weaning of newborn dairy calves. These milk replacers (MR) should accelerate the rumen development, determining early solid feed intake and leading to better performances in cattle. Previous research demonstrated that sodium butyrate supplementation in MR can affect both small intestine and rumen development. Also acetate and propionate showed similar properties, while only a few studies indicate some potential benefit of monoglycerides on gut functions. The present study is aimed to determine the effect of the supplementation of a blend containing short and medium chain fatty acids monoglycerides (SMCFA) in milk replacer on rumen papillae development and growth performances in weaning calves. Twenty bull calves (about 2 weeks old, weighing around 43kg) were randomly allocated into two groups: control (C) and treated (T). Besides MR and starter diet, the latter offered at libitum, T calves received 0.2% SMCFA in MR. Animals were slaughtered after 56 days from the beginning of the trial. No difference was found between groups either in growth performances or in mean number of papillae/cm(2) of mucosa, total surface of papillae (mm(2))/cm(2) of mucosa or papillary size. In both groups, the morphology of the rumen epithelium was typical of parakeratosis. The cells of the stratum spinosum were directly transformed into swollen, ovoid, still nucleated keratinocytes, particularly at the papillary tip, probably as a result of unphysiological osmolarities caused by high concentrate intake. Degenerated squamous horn cells covered the "balloon like" cells forming several layers, particularly in the places of the rumen mucosa more protected from an abrasive action of solid feed. This was more evident in C animals. The squamous cells covering the papillary tip showed cytoplasmic protrusion, representing remains of the attachment sites of desmosomes, which increased the total absorptive surface and were more numerous and higher in T compared to C animals. It might be hypothesized that SMCFA supplementation in MR could better regulate epithelial cell proliferation and probably have an "emollient effect" leading to an easier "peeling" that might increase efficiency for nutrient transport across the epithelium.

Localization of peripheral autonomic neurons innervating the boar urinary bladder trigone and neurochemical features of the sympathetic component.

The urinary bladder trigone (UBT) is a limited area through which the majority of vessels and nerve fibers penetrate into the urinary bladder and where nerve fibers and intramural neurons are more concentrated. We localized the extramural post-ganglionic autonomic neurons supplying the porcine UBT by means of retrograde tracing (Fast Blue, FB). Moreover, we investigated the phenotype of sympathetic trunk ganglion (STG) and caudal mesenteric ganglion (CMG) neurons positive to FB (FB+) by coupling retrograde tracing and double-labeling immunofluorescence methods. A mean number of 1845.1±259.3 FB+ neurons were localized bilaterally in the L1-S3 STG, which appeared as small pericarya (465.6±82.7 µm2) mainly localized along an edge of the ganglion. A large number (4287.5±1450.6) of small (476.1±103.9 µm2) FB+ neurons were localized mainly along a border of both CMG. The largest number (4793.3±1990.8) of FB+ neurons was observed in the pelvic plexus (PP), where labeled neurons were often clustered within different microganglia and had smaller soma cross-sectional area (374.9±85.4 µm2). STG and CMG FB+ neurons were immunoreactive (IR) for tyrosine hydroxylase (TH) (66±10.1% and 52.7±8.2%, respectively), dopamine beta-hydroxylase (DßH) (62±6.2% and 52±6.2%, respectively), neuropeptide Y (NPY) (59±8.2% and 65.8±7.3%, respectively), calcitonin-gene-related peptide (CGRP) (24.1±3.3% and 22.1±3.3%, respectively), substance P (SP) (21.6±2.4% and 37.7±7.5%, respectively), vasoactive intestinal polypeptide (VIP) (18.9±2.3% and 35.4±4.4%, respectively), neuronal nitric oxide synthase (nNOS) (15.3±2% and 32.9±7.7%, respectively), vesicular acetylcholine transporter (VAChT) (15±2% and 34.7±4.5%, respectively), leu-enkephalin (LENK) (14.3±7.1% and 25.9±8.9%, respectively), and somatostatin (SOM) (12.4±3% and 31.8±7.3%, respectively). UBT-projecting neurons were also surrounded by VAChT-, CGRP-, LENK-, and nNOS-IR fibers. The possible role of these neurons and fibers in the neural pathways of the UBT is discussed.

Neurochemical features of boar lumbosacral dorsal root ganglion neurons and characterization of sensory neurons innervating the urinary bladder trigone.

Porcine lumbosacral dorsal root ganglion (DRG) neurons were neurochemically characterized by using six neuronal markers: calcitonin gene-related peptide (CGRP), substance P (SP), neuronal nitric oxide synthase (nNOS), neurofilament 200kDa (NF200), transient receptor potential vanilloid 1 (TRPV1), and isolectin B4 (IB4) from Griffonia simplicifolia. In addition, the phenotype and cross-sectional area of DRG neurons innervating the urinary bladder trigone (UBT) were evaluated by coupling retrograde tracer technique and immunohistochemistry. Lumbar and sacral DRG neuronal subpopulations were immunoreactive (IR) for CGRP (30 ± 3% and 29 ± 3%, respectively), SP (26 ± 8% and 27 ± 12%, respectively), nNOS (21 ± 4% and 26 ± 7%, respectively), NF200 (75 ± 14% and 81 ± 7%, respectively), and TRPV1 (48 ± 13% and 43 ± 6%, respectively), and labeled for IB4 (56 ± 6% and 43 ± 10%, respectively). UBT sensory neurons, which were distributed from L2 to Ca1 DRG, had a segmental localization, showing their highest density in L4-L5 and S2-S4 DRG. Lumbar and sacral UBT sensory neurons expressed similar percentages of NF200 immunoreactivity (64 ± 33% and 58 ± 12%, respectively) but showed a significantly different immunoreactivity for CGRP, SP, nNOS, and TRPV1 (56 ± 9%, 39 ± 15%, 17 ± 13%, 62 ± 10% vs. 16 ± 6%, 16 ± 11%, 6 ± 1%, 45 ± 24%, respectively). Lumbar and sacral UBT sensory neurons also showed different IB4 labeling (67 ± 19% and 48 ± 16, respectively). Taken together, these data indicate that the lumbar and sacral pathways probably play different roles in sensory transmission from the UBT. The findings related to cell size also reinforced this hypothesis, because lumbar UBT sensory neurons were significantly larger than sacral ones (1,112 ± 624 µm(2) vs. 716 ± 421 µm(2) ).

Experimental study on the location of neurons associated with the first sacral sympathetic trunk ganglion of the pig.

The neurons associated with the left first sacral sympathetic trunk ganglion (STG S1), an autonomic ganglion particularly concerned in the innervation of the smooth and striated musculature associated with pelvic organs, were identified in the pig, using the non-trans-synaptic fluorescent retrograde neuronal tracer Fast Blue. The labelled neurons were located mostly ipsilaterally, in the intermediolateral nucleus of the spinal cord segments T10-L5, in the sympathetic trunk ganglia L3-Co1, in the caudal mesenteric ganglia, in the pelvic ganglia, and in the spinal ganglia T13-S4. Our results could indicate the existence of visceral neuronal circuits concerning the ganglia of the sympathetic trunk and the caudal mesenteric, pelvic and spinal ganglia with or without the intervention of the central nervous system, whose identification and preservation during surgical treatments could be helpful in reducing the risk of subsequent urinary and sexual disfunctions.

Double labelling immunohistochemical characterization of autonomic sympathetic neurons innervating the sow retractor clitoridis muscle.

Retrograde neuronal tracing and immunohistochemical methods were used to define the neurochemical content of sympathetic neurons projecting to the sow retractor clitoridis muscle (RCM). Differently from the other smooth muscles of genital organs, the RCM is an isolated muscle that is tonically contracted in the rest phase and relaxed in the active phase. This peculiarity makes it an interesting experimental model. The fluorescent tracer fast blue was injected into the RCM of three 50 kg subjects. After a one-week survival period, the ipsilateral paravertebral ganglion S1, that in a preliminary study showed the greatest number of cells projecting to the muscle, was collected from each animal. The co-existence of tyrosine hydroxylase with choline acetyltransferase, neuronal nitric oxide synthase, calcitonin gene-related peptide, leu-enkephalin, neuropeptide Y, substance P and vasoactive intestinal polypeptide was studied under a fluorescent microscope on cryostat sections. Tyrosine hydroxylase was present in about 58% of the neurons projecting to the muscle and was found to be co-localized with each of the other tested substances. Within fast blue-labelled cells negative to the adrenergic marker, small populations of neurons singularly containing each of the other enzymatic markers or peptides were also observed. The present study documents the complexity of the neurochemical interactions that regulate the activity of the smooth myocytes of the RCM and their vascular components.

Peripheral ganglia supplying the genital smooth musculature in the female pig: an experimental study.

The aim of the present study was to locate the sensory and autonomic ganglia innervating the female genital musculature in pigs. The retrograde neuronal tracers horseradish peroxidase (HRP) or fast blue (FB) were injected into the left retractor clitoridis muscle (RCM), which was treated as a typical model of the genital smooth musculature. Labelled cells were found in ipsilateral dorsal root ganglia S1-S4, in bilateral sympathetic paravertebral ganglia from L5-L6 or L6-L7 to S3 and in the left and right caudal mesenteric ganglion. In two of the five animals treated, presumably preganglionic parasympathetic cells were labelled in the ipsilateral intermediate grey substance of the segments S1-S2.

Further observations on the presence of ganglion cells in the oculomotor nerves of mammals and fish: number, origin and probable functions.

The oculomotor nerves (3rd, 4th and 6th) of some species of fish and mammals have been studied to establish the presence, number, true topography and probable functional role of the ganglion cells located along the trunk. The finding of typical pseudo-unipolar ganglion cells is always unpredictable and extremely variable, from an inter- and intra-specific point of view, in members of the two zoological classes studied.

On the presence of ganglion cells in the intracranial portion of the accessory nerve (XI cranial nerve) in some mammals.

The intracranial tract of the accessory nerve (XI cranial nerve) was studied in some mammals (equines, domestic and wild ruminants, pig, carnivores, rabbit, nutria, guinea pig, hamster, hedgehog). The specimens were embedded in paraffin or paraplast, the sections were stained with cresyl violet, haematoxylin and eosin, or submitted to argentic impregnation. Pseudounipolar ganglion cells were found in all the mammals examined, with the exception of the cat. The number of cells and their variability in the different species and subjects were related. The topography and morphology of the cells were described. This comparative study has demonstrated that the accessory nerve is not a entirely motor nerve, but it is a mixed, motor and sensitive, nerve. Nevertheless, we think further studies are necessary in order to establish the peripheral distribution, the central pathway and the functional role of the pseudounipolar neurons found in the intracranial tract of the accessory nerve.

Neurotoxic effects of 3-acetylpyridine (3-AP) in the hamster (mesocricetus auratus).

The 3-Acetylpyridine (3-AP) is a neurotoxic that determines a selective destruction of inferior olivary nucleus in the rat. However, in the other animals (mouse, rabbit, guinea-pig, quail and chicken) the effect could be less selective in the mentioned nucleus and constant in other areas of the central and peripheral nervous system. This investigation means to expand the study of these neurotoxic effects to the hamster, species not yet studied. The intraperitoneal administration of 3-AP, at a dosage of 65 mg/Kg, determines in this animal stridulous breathing, coughing and ataxy. Wide sites of the nervous system show, by microscopical observations, pictures of shrinking cells, vacuolization of the cytoplasm, reduction or loss of Nissl stain affinity and neuronal mutilation. The neurotoxic effects are mostly evident in the motor nuclei of the cerebral trunk and in the ventral horns of the spinal cord. However, the inferior olivary nucleus appears undamaged or sparely involved.

On the vegetative and sensitive innervation of the Retractor clitoridis muscle in some domestic animals.

The retractor clitoridis muscle originates from the coccygeal vertebrae in the cow, ewe, goat and mare, and from the anal musculature in the sow. It terminates at the base of the clitoris. In all the species considered, a vegetative innervation was found. This was represented by isolated or grouped ganglion cells. Nervous sensitive supply was also present. This was represented by Pacinian, Pacinian-like and Golgi-Mazzoni's corpuscles, and by Krauses's end bulbs. A notable difference was found in the amount and type of these receptors. They were numerous in the sow, ewe and goat, and rare in the cow and mare. Additionally, in the sow, ewe and goat, all the above mentioned receptors were found, while, in the cow and mare, only Pacinian and Pacinian-like corpuscles occurred. The morphology of these receptors was described and hypotheses were made concerning their probable functional role.

Comparative study of sensitive and vegetative innervation of external and internal anal sphincter muscles in different mammals.

Proprioceptive innervation of the external anal sphincter muscle and the organization of the vegetative and sensitive nerve components of the internal and sphincter muscle have been studied in different mammals. The findings of typical muscle spindles in the external anal sphincter muscle were constant in the pig, frequent in the goat and cow, rare in the sheep and horse and absent in the roe and rabbit. In the pig, muscle spindles were observed in the entire extension of the muscle, while in the sheep, goat, cow and horse, the receptors were found only in the cranial portion of the muscle. In all the species studied, the internal anal sphincter muscle had numerous ganglion cells, isolated or grouped, and rare Pacinian, Pacinian-like, and Golgi-Mazzoni corpuscles. Their functional role has been hypothesized.

The pyramidal tract of the hedgehog (Erinaceus europaeus) and its relationship with the olfactory bulb.

The pyramidal tract of the hedgehog has been investigated. The motor cerebral cortex was first located electrophysiologically and subsequently ablated. Ablation of the motor cortex did not apparently cause motor deficits. The cerebral hemisphere of the operated side, brain stem and spinal cord were removed for histological examination. The Nauta-Gygax and the horseradish peroxidase methods were employed to study the course and origin of the above mentioned tract. The pyramidal tract runs ipsilaterally in the cerebral peduncle, and does not go beyond the first cervical segments of the spinal cord (C3-C4); moreover, their fibers do not cross at spinal level. The close relation between this anatomical pathway and the olfactory bulb of the same side is also reported.

[Update on the presence of taste buds on the laryngeal surface of the epiglottis]. / Ancora sulla presenza di calici gustativi nella faccia laringea dell'epiglottide.

Previous investigations have ascertained, according to the results obtained by Bradley et al. (1980) in the sheep, that the laryngeal surface of the epiglottis of goat and bovine is always provided with numerous taste buds. These observations have verified in other ruminant species, as the moufflon and the buffalo, the validity of the above-named datum and have ascertained that it is always inconstant in the other animal species considered (wild boar, coypu). These taste buds show a typical structure (diameter of the outer taste pore varying from 2.7 to 4.2 micron, width of the chemoreceptors varying from 30 to 60 micron and length from 27.5 to 57.5 micron). Moreover, the normal structure of the above-named taste buds is also testified by the arrangement of their innervation and particularly by the integrity of the synaptic contacts. The results of the present research have permitted a critical and more severe examination of the probable functional role of those laryngeal receptors. In fact, in the ruminants they may protect the deep airways precluding to food particles the larynx in the phase of food regurgitation.

[Neuromuscular spindles of the extrinsic ocular musculature of the moufflon]. / I fusi neuromuscolari nella muscolatura estrinseca oculare del Muflone.

Proprioceptive innervation of moufflon extrinsic ocular musculature and m. levator palpebrae superioris was studied. Muscle spindles and Golgi tendon organs were found. The first ones are usually between 1st and 2nd order muscle fascicles. The muscle spindles are highly represented in the extrinsic ocular muscles, but less numerous in m. levator palpebrae superioris. Their number varies according to muscles and individuals. In the same subject, also the ratio between the number of the muscle spindles found in m. rectus dorsalis and that of m. levator palpebrae superioris was examined. Besides, the histological structure of the intrafusal fascicles was investigated. Particular attention was devoted to the nerve supply of the muscle spindle. By means of impregnating methods, sensory and motor endings were identified. Primary and secondary sensory endings only in a few cases showed their usual pattern: motor fibres can end in form of plates or trails. Golgi tendon organs were observed between the tendon and the muscular tissue and are always less numerous.

Coccygeus and levator ani muscles in the rabbit: morphology and proprioceptive innervation.

The Authors have studied the morphological features and the proprioceptive nervous component in the coccygeus and levator ani muscles of the rabbit, using Ruffini's and Barker-Ip's impregnations. The coccygeus muscle originates from ischiatic spine and inserts on the last three sacral vertebrae and on the first four or five caudal vertebrae. The levator ani muscle originates from the ischiatic spine and the coccygeus muscle aponeurosis and inserts directly on the caudal vertebrae 3-5 or 4-6. The proprioceptive innervation in both muscles is constituted by muscle spindles and Golgi tendon organs with a typical structure. Muscle spindles are more numerous than Golgi tendon organs and the spindle density is higher in the levator ani muscle.

Course and termination of the pyramidal tract in the pig.

To study the pyramidal tract in the pig, the motor cerebral cortex of one side was defined electrophysiologically and subsequently excised. The animals operated were killed after 7, 11 and 14 days, and the cerebral hemisphere of the operated side, brain stem and spinal cord were removed for histological examination. The pyramidal tract proved to run ipsilaterally as far as the oral extremity of the 12th cranial nerve nucleus. The decussation, which exhausted itself almost completely at the level of the rostral extremity of the 1st cervical metamere, started here. After the limit just mentioned only rare isolated fibres were visible. Along its course, the pyramidal tract sent a small number of axons to the ipsilateral and contralateral nucleus of the 7th cranial nerve, while the fibres running from the opposite side to the reticular formation and to the hypoglossal nerve nucleus, cuneatus, gracilis and trigeminal spinal tract nuclei were more numerous.

Macroscopic organization and sensitive innervation of the tendinous intersection and the lacertus fibrosus of the biceps brachii muscle in the ass and horse.

The most developed and organized lamina running trough the biceps brachii muscle belly forms a well-marked tendinous intersection connecting the proximal tendon with the distal one. Moreover, the lacertus fibrosus arises from this lamina close to the distal tendon and blends with the fascia of the forearm and joins the extensor carpi radialis muscle. The nerve supply for the biceps tendinous intersection arises from the intramuscular rami of the musculo-cutaneous nerve, whereas the lacertus fibrosus is provided with some cutaneous rami of the same nerve. The biceps brachii tendinous intersection and lacertus fibrosus are provided with free and encapsulated nervous endings. The encapsulated corpuscles are represented by rare Pacini, Pacini-like and Ruffini receptors and by numerous Golgi tendon-organs. All these receptors have been studied by a topographical point of view and the role played by the proprioceptors in synchronizing shoulder, elbow and carpal joint movements both in the flexion and the extension of the limb, has been hypothesized.