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Determination of Immunobiological Reaction in Sheep
Chlamydiosis
Navruzov N.I.,
junior researcher.,
Sayfidinov B.F.,
intern researcher.,
Aktamov U.B.,
intern researcher.
Veterinary Scientific Research Institute
------------------------------------------------------***-------------------------------------------------------
Abstract:
The article reports on the stability of the immune system in the div when using
the GOA formalin vaccine against sheep chlamydiosis.
Key words:
Chlamydia, immunoglobulin, vaccine, immunity, immunophone, antigen,
microorganism, receptor, serotype.
Relevance of the research.
Chlamydiosis in small and big horned animals is well
documented to inflict significant economic harm to all farms in our nation. Investigations have
shown that chlamydia is responsible for an average of 12 percent of abortions in animals. Up to
50% of miscarriages in farm animals are caused by chlamydia, claim I.I. Nosov and A.A. Volkova.
The care of sick animals and the efforts to stop the sickness cost a lot of money.
According to the literature, chlamydia prevalence rates for small horned animals were 18-
43.6 in the USA, 9-21.6 in Canada, 16-24.8 in the Netherlands, 18-57 in France, 14-46.4 in
England, 6-26.2 in Australia, and 3-34 percent in Israel.
Chlamydia is a contagious, enzootic infectious illness that causes inflammation of the
lungs, abortion in the second part of the cervix, incompetent lambs and calves (usually in young
animals), and inflammation of the placenta, especially the cotyledons. (pneumonia).
Chlamydia abortus ovis, a member of the Chlamydiacea family and Chlamydiacea psittaci
genus, is the responsible party. Chlamydia is a parasite that may grow up to 250–300 nm in
size. They are bacteria with thick cell walls that carry DNA and RNA.
The chlamydia-causing agent has a complicated antigen structure with three antigenic
centers that are unique to the genus, species, and serogroup. Its gender is lipopolysaccharide
because, like gram-negative bacteria, it has a thermostable cell wall. The binding epitope, which
establishes the specificity of the genus and includes a unique receptor found in carbohydrates,
and an oligosaccharide molecule with three monomers are the two components that define
antigenicity. (G.A. Dmitriev et al., 1999). The particular location of cysteine-rich amino acids in
the protein membrane of species-specific determinants varies between antigen serotypes.
Purpose of the research.
The key criteria of our experiment to assess the efficacy of the
vaccination is stability of the impact of the GOA formalin vaccine against sheep chlamydiosis on
INTERNATIONAL JOURNAL ON ORANGE TECHNOLOGY
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the immune system using IgM and IgG test kits created by JV "UNIGEN."
Research object and methods.
Research was conducted in VITI's Microbiology, Regional
Diagnostics, and Young Livestock Disease Research Laboratories as well as under production
circumstances at the livestock complex in the Kashkadarya region's Dehqonabad district called
"Karakolchilik Shirkati named after M. Ibragimov."
The quantity of immunoglobulins and how they affected the infections affected the div's
ability to fight against germs. Farm animals almost never have immunoglobulin-E or
immunoglobulin-D identified in them. (F.J.Bourne et al. 1978). The first step of immunological
responses is marked by the appearance of IgM from macroglobulins. The primary
immunoglobulin in blood serum is IgG, which comes in two varieties: Ig–G
1
and Ig–G
2
. The
primary cellular components of the div, in addition to immunoglobulins, are macrophages
(monocytes), as well as active T and V lymphocytes, which guarantee the div's resistance
against pathogens and viruses. (Green S.A., Albulov A.I., Ruban E.A., Green A.V. .). The
morphological and pathological status of the div's tissues and cells is negatively impacted by
the antibiotics used to treat the disease. It is important to remember that polyclonal activation
syndromes are what lead to false positive outcomes in such enzymatic and sequential
processes. The creation of specific defensive protein enzymes against diverse foreign antigens
entering the div is simultaneously stimulated by superantigens, which are present in the
individual (ontogonistic) phase of the animal div, and V cells respond in a particular manner.
In actuality, these activities manifest as a simultaneous, non-specific rise in the titer of antigens
to several infections. According to literature sources, technical flaws in the production of the
response as well as immunodeficiency circumstances may be to blame for false negative
findings in the detection of antigens. A specific apparatus (colorimeter) was used to measure
the color intensity of the liquid in the tablet wells after the reaction had halted, and a special
gadget was utilized to compute the results. The control samples' optical densities were
compared, and the analysis's findings underwent mathematical processing. It was determined
that the sample contained more particular chlamydial antibodies the greater the optical density
in this well.
Antigen was preadsorbed on the well walls of 96-well polystyrene tablets for IFT. The
well of the tablet was filled with the blood serum that would be evaluated. In this instance, the
homologous antibodies join after attaching to the previously adsorbed antigen. During washing,
chlamydia antibodies that are not bound are eliminated. Chlamydia antibodies were then
introduced to the well along with enzyme-labeled antibodies against rabbit or other animal
immunoglobulins. If any chlamydial antibodies are found in the blood serum under
examination, they act as antigens at this point and mix with chlamydial antibodies found for the
enzyme. After washing, a chromogenic (coloring) material was applied, allowing for the
possibility of accounting for the reaction on the growth of staining in the wells. Since staining
intensity is inversely correlated with enzyme concentration, chlamydia antidiv concentration
is likewise inversely correlated with enzyme concentration. (they are quantitatively
equivalent). The optical density of the liquid in the wells was measured and compared to the
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optical density of the control sample to determine the antidiv concentration per unit volume.
Results were calculated using optical density units. For taking IFT findings, levels of normal
indicators, and pathological indicators into consideration, each test system has its own
indicators. Results of immunoenzyme analysis are based on them.
IFT was conducted using "Socorex" dispensers, ELx405 microplate cleaning apparatus, and
ELx808 microplate automated analyzers. Using a computer and the Bio-Tek KC4TM software,
the reaction's findings were electronically (and hence electronically) interpreted.
In a test to determine the preventative efficacy of IgM and IgG test kits created by
"UNIGEN" and "XEMA" LLC, serological and immunological responses in the div of sheep that
had received the vaccination were examined.
Ten sheep in experimental group I received two subcutaneous injections of the
"emulsified vaccine against chlamydiosis."
Only one of the ten heads in the II comparative group received the "Chlamydiosis vaccine."
Group III (10 heads) served as the control group, receiving no medication. Based on the
farm veterinarian's anamnesis data and taking into consideration the fact that the lambs that
miscarried and gave birth the previous year were not viable, the sheep chosen for the study
were decided.
Results.
Based on the short-term detection of the response to the inciting antigen or the
particular antidiv generated against it, we investigated immunoenzymatic analysis (IFT). In
spite of the fact that the serological (KBR) method can distinguish between infected and
vaccinated animals in the case of small horned animals immunized against chlamydia in the
experiment, we used it to find that it is simpler, quicker, and more practical to diagnose than
the immunological method. (1 -table).
Optical density values Table 1
Optical Density Values
1
2
3
4
5
6
7
8
9
10
11
12
A
1,413 0,062 0,867 0,245
B
1,451 0,069 0,962
C
0,257 0,136 0,164
D
0,224 0,228 0,135
E
0,096 0,923 1,899
F
0,092 1,926 0,892
G
1,517 0,038 0,055
H
1,578 0,052 0,034
The samples in the first pair (A1 and V1) are the optical density standards for negative
samples, while the samples in the next pair are for positive blood samples, as can be seen from
the findings of the table. (S1 and D1). Blood samples from cells G1, H1, F2, and E3 were found
to be positive in the examination of 21 examined blood samples, whereas blood samples from
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cells E2, A3, and B3 were determined to be suspicious.
Based on the high specificity and sensitivity of "antigen-antidiv" immunological
responses, enzyme immunoassay is a laboratory experiment. IFT is made up of two distinct
parts: immunological and enzymatic processes. The immunological response (bacteria and
viral molecules) acted as an antigen and a binding site for antibodies.
Additionally, the outcomes of the immunological reaction may be observed and quantified
thanks to the enzymatic reaction. In order to manage the epizootological situation and ascertain
the general immunophone in the farm of "Karakolchilik shirkat named after M. Ibragimov" LLC,
Kashkadarya area, Dehqonabad district, immunoenzymatic analysis (IFT, ELISA) reaction was
utilized as an immunobiological approach. Prior to starting the reaction, precautions were
taken to assure compliance with the rules and guidelines for biological safety in the laboratory.
Detecting IgG-specific antibodies against chlamydia in the blood sera of cattle and small
horned animals using immunoenzymatic analysis (IFT or ELISA). Chlamydia IgG-IFA was
created in collaboration with the Veterinary Institute of Veterinary Medicine and the joint
ventures of "UNIGEN" and "XEMA" LLC. It is an IgG antidiv against the chlamydia-causing
agent. The test system was used to perform "Set of reagents for detection with" ("Nabor
reagentov dlya immunofermentnogo vyyavleniya IgG antitel k vozbuditelyu chlamydiosis
krupnogo i melkogo hornatogo skota"). (Figures 1, 2). and were carried out in accordance with
the standard guidelines for carrying out the IFT response.
Figure 1 IgM-
immunoglobulin
Figure 2 IgG-
immunoglobulin
Table 2
Immunological analysis of chlamydiosis vaccine.
Groups
Number of
animals
Types of analysis
C-reactive protein
(normally 0.1-0.3 mg/l)
IgM (normally
0.4-2.3 mg/l)
IgG
(normally 7-
16 mg/l)
Experimenta
l group I
10
0,32±0,025
2,8±0,23
17,1±0,96
Comparative
group II
10
0,287±0,015
2,04±0,143
16±1,144
Control
group III
10
0,108±0,0058
0,286±0,022
7,3±0,46
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In experimental group I, it was discovered that the level of C-reactive protein was 1.55
times greater than the average. It was discovered to be 1.13 times greater than the first group
II, which was in the comparative experimental group II at the standard level. It was discovered
that experimental group I was 1.69 times higher than the norm than the control group, and
group II was 1.21 times higher when assessing the course of the disease in a chronic condition
according to the change of IgG. IgM and IgG levels were found to be particularly high in the first
group, with just a slight difference from the II group and a significantly greater effect from the
III group.
Conclusions:
1. From February to May, animal farms in the Samarkand area tested positive for 8.2
percent of chlamydia; farms in the Kashkadarya region tested positive for 6.9 percent of
chlamydia.
2. Despite the excellent sensitivity and accuracy of the responses in both cases when
diagnosing chlamydiosis by serological (KBR) and immunological (IFT) approaches, it was
discovered that immunoenzymatic analysis was simple to perform.
3. The chlamydiosis-causing agent's cultural, morphological, tinctorial, virulence,
biochemical, and pathogenic properties were detected in several cattle farms with a focus on
breeding in the Kashkadarya and Samarkand areas.
4. Based on the sensitivity of chlamydia to antibiotics, it was shown that erythromycin is
less sensitive than oleandomycin, doxilox, teliosin, oxacillin, and doxilox.
References
1.
Розанов
Н.И.
“Микробиологическая
диагностика
заболеваний
сельскохозяйственных животных”. Москва, Государственное издательство
сельскохозяйственной литературы, 1952, 508 с.
2.
Сидоров М.А., Скородумов Д.И., Федотов В.Б. “Определитель зоопатогенных
микроорганизмов”. Москва, “Колос”, 1995. 319 с.
3.
Кисленько, В.Н. Ветеринарная микробиология и иммунология // М.: Колос, 2007. -
224 с.
4.
Колычев Н.М., Кисленько В.Н., Суворина О.С. Частная микробиология // -М. : Колос С,
2007. 215ст.
2013 год, кандидат
наук Тюлькова Лариса Сергеевна.
6.
Hokinson R.G., Griffiths P.C., Rankin S.E.S. Towards ad: ferential polymerase chain reaction
test for Chlamydia psittaci. Vet. Tec., 1991, 128;-с. 381-382.
7.
Kaltenboeck B. Structures of and allelic diversity and relationships among the major outer
membrane protein (ompl) genes of the Chlamydia species. J. Back. 1993 V. 175.- P.478-
502.
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8.
Самуйленка А.Я., В.Н., Сюрин Е.С. Воронин // Инфекционная патология животных:
Том V – Хламидиозы – Москва 2003. – С.10-12.
9.
Гнездилова Л.А. Эпизоотологическая характеристика, диагностика, клинические
проявления хламидиоза овец / Л.А. Гнездилова, М.А. Викулова // Сб. науч. тр. М.
2006. - Ч. 2 – С. 9-11.
10.
Митрофанов П.И., А.А.Сидорчук., Л.А.Гнездилова. // Хламидиозы животных Москва
2006. – С. 45-46.
