CC BY
20«COyyOMUM-JMTMaiL» #794)), 2©21 / VETERINARY science
7
Veterinary science
Horalskyi L.P., Kovalchuk O.M.,
Polissya National University, Zhytomyr, Ukraine
Gutyj B.V.,
Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv, Ukraine
Sokulskyi I.M.
Polissya National University, Zhytomyr, Ukraine DOI: 10.24412/2520-6990-2021-794-7-12 PATHOMORPHOLOGICAL FEATURES OF THE PANCREAS IN MATURE CATS WITH ACUTE
PANCREATITIS
Abstract.
The aim of the study was to investigate the pathomorphology of the pancreas of adult cats with acute pancreatitis. Histological examinations of the pancreas were performed in the laboratory at the Department of Anatomy and Histology of Polissya National University.
Histological sections were stained with hematoxylin and eosin and histomorphometric analysis of parts (exo-and endocrine) of the pancreas was performed for examination microscopy to detect histological changes in cats with acute pancreatitis. The obtained morphometric data indicate an increase in sick animals in 1.12 times the parenchyma of the exocrine part of the gland, which was 48257.09 ± 135.06 pm2 (97.89 ± 0.04%). The endocrine part, respectively, occupied 6771.88 ± 274.06 p.m2 area (3.89 ± 0.03%.). In the control group of animals, the corresponding values were 42989.03 ± 141.05 pm2 (96.11 ± 0.05%) and 6811.44 ± 312.04 (3.89 ± 0.03%).
Characteristic pathohistological changes in the gland were manifested by edema of the intercinary space and an increase (p <0.05), 1.27 times the proportion of its stromal component, which was 22.47 ± 0.25%, and in the control - 17.69 ± 0.14% respectively.
Keywords: morphology, histostructure, islets of Langerhans, lysis of nuclei, proliferation of nuclei, volume ofpancreatocytes.
Introduction.
The pancreas is one of the few organs that, due to the combination of external secretory and endocrine functions, participates in almost all physiological processes, from digestion to adaptation processes, including the maintenance of homeostasis of the whole organism [3, 4]. The activity of the body is extremely important for the regulation of carbohydrate, protein and fat metabolism in animal and human tissues [11].
It was found that diseases of the digestive system in animals are about 45% out of the total number of non-communicable diseases, among which in the first place is inflammation of the pancreas [12].
However, the data of morphological studies on the pathology of the pancreas in animals are ambiguous, and the results of histological studies in sick cats are given only superficially.
There is no doubt that pancreatitis in domestic animals, including cats, remains one of the most common diseases and insufficiently clarified causes of morbidity and mortality of animals of this species [6, 8].
Pancreatitis is an inflammatory-dystrophic disease of the glandular tissue of the pancreas that occurs due to premature activation of proenzymes in acinar cells and occurs with impaired patency of its ducts [7]. This pathology can be both acute and chronic, bacterial and nonbacterial in nature [10]. It depends on the degree of manifestation of its atrophic processes associated with fibrosis in the parenchyma of the pancreas [13]. However, there may be a severe form of the disease, which is accompanied by the development of pancreatic necrosis and systemic complications [1]. Any form of
pancreatitis can occur with mild symptoms or manifested by nonspecific clinical signs [10].
The acute form of pancreatitis is accompanied with edema and necrosis, with hypoperfusion and thrombosis, which in turn can exacerbate pancreatic necrosis [5].
At the same time acute pancreatitis can turn into chronic, which in turn may be accompanied by exocrine insufficiency of the pancreas.
The most common signs of pancreatitis in cats are low body temperature, anorexia, dehydration, jaundice is less common [9].
Therefore, given the importance of the function of the pancreas not only as an organ of the digestive system, but also one that determines the general condition of the whole organism, it is important to study the pathomorphological features of the gland in cats with acute pancreatitis.
Material and methods.
In conducting research, the basic rules of good laboratory practice GLP (1981), the provisions of the "General Ethical Principles of Animal Experiments", adopted by the First National Congress of Bioethics (Kyiv, 2001) and the requirements for the Rules of experimental use animals ".
The material for the pathomorphological study was cats that died from pancreatitis diagnosed on the basis of clinical, biochemical studies and ultrasound (n = 6).
Histological examination of the pancreas in normal and in pathology was performed in the pathomor-phological laboratory of the Department of Anatomy
s
VETERINARY SCIENCE / «©OLiyOMUM-JMTMaiL» 2©2U
and Histology, Faculty of Veterinary Medicine, Results.
Polissya National University. The pancreas in cats is externally covered with a
To study the morphology of cells and tissues, and thin connective tissue capsule. The parenchyma of the
morphometric study used staining of histocuts with he- gland is divided into lobes, between which are the septa matoxylin Ehrlich, Karats and eosin and the method of (Fig. 1), which are connective tissue strands in which
Van Gizon [2]. blood vessels, nerves and excretory ducts.
Microphotography of histological specimens was performed using a camcorder CAM V200 mounted in a microscope Micros MC-50.
Fig. 1. A fragment of the microscopic structure of the exocrine part of the pancreas of the control group of cats: 1 - acinus; 2 - interparticle connective tissue. Hematoxylin Karatsi and eosin. x 120.
Morphologically, it is noted that the largest part of the gland is the exocrine part (Fig. 3), which according to morphometric studies of the pancreas in clinically healthy adult cats is 42989.03 ± 141.05 ^m2 per unit area of the gland in optical examination (approx. 16, about 8) or (96.11 ± 0.05%). It consists of terminal divisions - pancreatic acinuses with a diameter of 29.89 ± 0.15 ^m and excretory ducts.
The volume of pancreatocytes and their nuclei in control animals is 425.05 ± 17.11 and 30.92 ± 1.60 ^m3, respectively. The nuclear-cytoplasmic ratio of pancreatocytes is 0.12 ± 0.003 um. from
The acinus of the gland is formed by secretory (ac-inous) cells, which are known to produce digestive en-
zymes: lipase, amylase, trypsin, chymotrypsin, nucleases, etc. [11]. On histological examination, such cells are mostly triangular in shape, the nuclei are rounded, located almost in the center of the cells, the cytoplasm of acinar cells is stained unevenly. Thus, the basal part of such cells is stained basophilically blue, which is called a homogeneous zone. The apical part has an ox-yphilic color, ie painted red - zymogenic zone. This color structure is due to the uneven distribution of organelles in the relevant parts of the pancreatocytes. In addition to cells, the exocrine part contains plug-in and intraparticle excretory ducts, which are lined with a single layer of epithelium.
«COyyOMUM-JMTMaiL» #794)), 2©21 / VETERINARY SCIENCE
9
# .
- -i ... ¿p»
«• »
1
f
5
<
-»If J*'"/, f
' *,
' r »
* » »
m
4 4
* . L«. _
• > . • s ' * ;
t V
;V»'
\ > ' 3
« // —\
'W S
- j&fEËJÊÊM
— /LJ^: - Or 0 * ^ i
3?T - v v.-, i
Fig. 3 Fragment of the microscopic structure of the pancreas of the control group of cats: 1 - exocrine part; 2 - nuclei of pancreatocytes; 3 - interparticle connective tissue; 4 - interstitial connective tissue 5 - vessel. Hematoxylin Karatsi and eosin. x 400.
The endocrine part of the gland, whose area is 6811.44 ± 312.04 (3.89%) per conventional unit area in the field of view of the microscope (approx. 16, vol. 8) is formed by pancreatic islets (islets of Langerhans), which are located between the acinuses (Fig. 3). Such islets consist of insulocytes, around which are fenestrated capillaries with pericapillary space. Small cells - insulocytes had a light cytoplasm. It is known that there are the following cell types: acidophilic - A-
cells, basophilic - B-cells, dendritic - D-cells and PP-cells. Histologically, it is noted that in the center of the islets of Langerhars the largest mass is formed by B-cells containing a significant amount of heterochroma-tin.
Sectional macroscopic studies showed that in cats with acute pancreatitis at autopsy, the pancreas was dark red, hyperemic, enlarged, swollen, with localized hemorrhage.
I-
Fig. 4. Microscopic structure of the pancreas of cats with acute pancreatitis: 1 - interstitial edema; 2 - discom-plexation of acinuses; 3 - pancreatocytes; 4 - nuclei ofpancreatocytes. Hematoxylin Karatsi and eosin. x 400.
Histological examination of the histostructure of the exocrine gland in acute pancreatitis in comparison with the control group of animals showed that the venous vessels and arteries were dilated, overflowing
with erythrocytes and single neutrophils. Isolated hemorrhages in loose fibrous connective tissue or in the lumen of the excretory ducts were noted. In addition, in addition to neutrophils, macrophages and lymphocytes were found in the gland stroma. In some cases, in the
10
VETERINARY SCIENCE / «©SyyOMUM-JOUTMaiL» #7(94), 2©2U
field of view of the microscope there was an expansion of the intercinary space, which indicated the initial stage of acute inflammation. Morphometric studies showed an increase in the proportion of stromal component (p<0.05) in 1.27 times, which was 22.47 ± 0.25% relative to animals in the control group - 17.69 ± 0.14%, respectively.
At the same time, morphological examination of the pancreas in cats with acute pancreatitis showed an increase of 1.12 times (p<0.001) in the exocrine parenchyma, which was 48257.09 ± 135.06 ^m2 or (97.89 ± 0.04%). Therefore, the mo rplio mc t ri cpa ra mc t c rs of the
m
endocrine part were reduced by 39.56 ^m2 and amounted to 6771.88 ± 274.06 ^m2. Such morphological parameters in control animals were 42989.03 ± 141.05 ^m2 (96.11 ± 0.05%) and 6811.44 ± 312.04 ^m2 (3.89 ± 0.03%), respectively.
Disorganization of the structure of individual exo-crinocytes was also noted. The latter were in a state of uneven swelling of the zymogenic zone, and a clear deformation of the homogeneous zone. The nuclei of such exocrinocytes moved to the basal part of the cell. Some exocrinocytes were in the stage of necrotic changes. There was a pronounced interstitial edema (Fig. 4).
t*
2
1
3
4
Fig. 5. Microscopic structure of the pancreas of cats with acute pancreatitis: 1 - plethora of the interciliary space; 2 - neutrophilic leukocytes; 3 - karyopyknosis of nuclei; 4 - smoothing the contours of the boundaries of
exocrine cells. Hematoxylin Karatsi and eosin. x 400.
In the analysis of the histrostructure of the acinus of the gland, found mostly destructive changes of various types. Thus, most noted a clear loss of eosinophilia of the cytoplasm, expansion of the perinuclear space, changes in the nuclei (compaction), the polarity of such cells was violated. Neutrophilic leukocytes were detected in such acinuses. In addition, the boundaries of exocrine cells lost their contours (Fig. 5). Focal hemorrhages were found in the parenchyma of the exocrine part. The vessels were dilated and filled with blood.
In addition, significant necrosis of the exocrine part of varying degrees was detected. In some parts of the parenchyma of the pancreas necrosis completely affects several lobes together with the connective tissue layer, in other parts of the parenchyma - only a small part of the lobes. Some acinuses lost their characteristic shape and did not have a clear structure (Fig. 6).
«COyyOMUM-JMTMaiL» #7(94), mm / VETERINARY science
11
Jf
r
jH
»
J
M jftj
A i
I
M
r
ft'«
4 «
1
ct
Fig. 6. Microscopic structure of the pancreas of cats with acute pancreatitis: 1 - necrosis ofpancreatocytes; 2 -
karyolysis. Hematoxylin Karatsi and eosin. x 400.
Microscopic examination of histological sections in the endocrine part showed that the contours of the islets of the pancreas were blurred, in a state of atrophy, in isolated cases, hyperplasia and sclerosis were observed in the area of histo sections. The number of islet cells in sick cats decreased significantly with the comparison of the control group. The walls of the vessels of
the islets were swollen, showed dilatation with intensive filling of the capillaries of the islets of Langerhans (Fig. 7), a slight stasis in the microcirculatory tract. Fragmentary necrosis of A-, D-, PP-, B-cells located on the periphery of the islets was noted.
Fig. 7. Microscopic structure of the endocrine part of the pancreas of cats with acute pancreatitis: 1 - destruction of cells of the islets of Langerhans; 2 - focal hemorrhages in the endocrine parenchyma. Hematoxylin Karatsi and eosin. x 400.
Morphometric examination of the diameter of the islets of Langerhans of the pancreas of sick cats of all studied groups revealed significant differences. Thus, the diameter of the acinuses and islets of Langerhans significantly (p <0.001) increased by 1.14 and 1.15 times and was equal to 31.84 ± 0.04 and 71.82 ± 2.1
^m, respectively. The corresponding values in clinically healthy cats were 27.88 ± 0.14 and 62.15 ± 2.45 ^m.
Conclusion.
1. Our histological morphometric and statistical methods of histoarchitectonics of pancreatic tissue of cats allows us to assess the degree of its morphological
1
2
12
VETERINARY SCIENCE / «©OLyOMUM-JOUTMaiL» 2©2fl
structure in normal and in terms of pathology - acute pancreatitis.
2. The histological structure of pancreatic drugs indicates cyto- and morphometric changes in the exocrine and endocrine parts. The results of morphometric studies of the exocrine part indicate a tendency to increase (p <0.001) in 1.12 times the exocrine parenchyma of cats, equal to 48257.09 ± 135.06 ^m2 or 97.89 ± 0.04%. The endocrine part of the gland, in contrast, was reduced by 39.56 ^m2 times and amounted to 6771.88 ± 274.06 ^m2. Such morphological parameters in control animals were 42989.03 ± 141.05 ^m2 -96.11 ± 0.05%% and 6811.44 ± 312.04 ^m2 - 3.89 ± 0.03%.
References
1. Andreeva, S.D., & Kirillovyh, A.S. (2013). Ul'tramikroskopicheskoe stroenie mikrocirkuljator-nogo rusla podzheludochnoj zhelezy pri ostrom destruktivnom pankreatite. Vestnik veterinarii. Materi-aly Mezhdunarodnoj nauchno-prakticheskoj internet-konferencii, posvjashhènnoj 65 letiju Stavropol'skogo GAU, «Sovremennye tendencii v veterinarnoj medicine». Stavropol': SGAU, 16, 30-33.
2. Horalskyi, L.P., Khomych, V.T., & Kononskyi, O.I. (2019). Osnovy histolohichnoi tekhniky i mor-fofunktsionalni metody doslidzhennia u normi ta pry patolohii: navch. posib. Zhytomyr: Polissia, 288.
3. Didenko, I.S. (2017). Zminy histolohichnoi struktury pidshlunkovoi zalozy za umov aloksanovoi hiperhlikemii u shchuriv u vikovomu aspekti. Ukrain-skyi zhurnal medytsyny, biolohii ta sportu, 6(8), 17-20.
4. Dubych, I.M. (2008). Histomorfolohiia ta orha-nometrychni pokaznyky pidshlunkovoi zalozy tsutse-niat. Nauk. visnyk Lvivskoho natsionalnoho univer-sytetu veterynarnoi medytsyny ta biotekhnolohii im. S.Z. Hzhytskoho. Lviv, 10(3(38)), 73-76.
UDC 619.09.636:678.048:615.916:631.842:636.4
5. Zinenko, D.Yu., & Berehovenko, I.M. (2008). Mikrotsyrkuliatorni y patomorfolohichni zminy u rozvytku eksperymentalnoho hostroho pankreatytu u shchuriv. Morfolohiia, 2(1), 33-40.
6. Tymoshenko, O.P. (2007). Informatyvnist son-ohrafichnoho doslidzhennia u diahnostytsi zakhvori-uvan pidshlunkovoi zalozy sobak. Visnyk Bi-lotserkivskoho derzh. ahrarnoho un-tu: Zb. nauk. prats. Bila Tserkva, 44, 101-103.
7. Katten, I. (2013). Sochetaniepankreatita s hol-angiogepatitom i vospalitel'nym zabolevaniem kishechnika (triadit) u koshek. Fokus, 23(2), 4-10.
8. Kravchenko, S.O., & Bobrova, V.V. (2018). Ul-trasonohrafichni zminy za hostrykh ta khronichnykh zapalen pidshlunkovoi zalozy u sviiskykh kotiv. Visnyk Poltavskoi derzhavnoi ahrarnoi akademii, 1, 138-142.
9. Ksenulis, P., & Shtainer, Y. (2009). Pankreatyt u koshek. Fokus, 19(2), 11-20.
10. Pankreatyt sobak. Monohrafiia: L.P. Ho-ralskyi, O.P. Tymoshenko, B.V. Borysevych ta in; Za redaktsiieiu L. P. Horalskoho. Zhytomyr: «Polissia», 215.
11. Gushhin, Ja.I. i dr. (2018). Sravnitel'naja mor-fologija podzheludochnoj zhelezy jeksperimental'nyh zhivotnyh i cheloveka. Laboratornye zhivotnye dlja nauchnyh issledovanij. Nauchno-prakticheskij zhurnal. Nauchno-prakticheskij zhurnal, 3, 33-48. doi: 10.29296/2618723X-2018-03-04
12. Tymoshenko, O.P., & Busel, Yu.M. (2009). Efektyvnist kompleksnoi diahnostyky pankreatytu v sobak, pidtverdzhena morfolohichnymy doslidzhen-niamy. Visnyk Poltavskoi derzh. ahrar. akad., 1, 87-93.
13. Akol, K.G., Washabau, R.J., Saunders, H.M., & Hendrick, M.J. (1993). Acute pancreatitis in cats with hepatic lipidosis. Journal of Veterinary internal Medicine, 7, 205-209.
Leskiv Kh Ya., Gufriy D.F., Gutyj B.V.,
Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies,
Pekarska, Str., 50, Lviv, 79010, Ukraine
Khalak V. I.
State Institution Institute of grain crops of NAAS, V. Vernadsky Str., 14, Dnipro, 49027, Ukraine
Hariv I.I., Martyshuk T. V., Guta Z.A.
Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies,
Pekarska, Str., 50, Lviv, 79010, Ukraine DOI: 10.24412/2520-6990-2021-794-12-15 THE EFFECT OF METHIPHENE, METHIONINE, AND PHENARONE ON THE HUMORAL PART OF THE IMMUNE SYSTEM OF PIGLETS IN EXPERIMENTAL CHRONIC NITRATE-NITRITE
TOXICOSIS
Abstract.
The article describes the dynamics of indicators of the state of humoral immunity ofpiglets in experimental nitrate-nitrite toxicosis. The subjects were 20 large white piglets three months old. The research was conducted at the Komarnivsky National Research Center of the Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies. According to the method of analog groups, one control and three experimental were formed. Feeding sodium nitrate to piglets at a dose of 0.3 g NO3'/kg. b.w. contributes to the development of chronic nitritenitrate toxicosis. Nitrates in a toxic dose for 90 days reduce the protective properties of the body. Under nitrate load, feeding pigsphenarone, methyphene, and methionine had a positive effect on piglets' humoral immunity, manifested by the high antimicrobial activity of serum.
