Vol.1 (2), pp. 9-15, December 2016

International Standard Journal Number ISJN: A4372-2601

Article Number: DRJA22847051

Copyright © 2016

Author(s) retain the copyright of this article

Direct Research Journal of Veterinary Medicine and Animal Science (DRJVMAS)

http://directresearchpublisher.org/journal/drjvmas

Research Paper

Fetal differentiation of the spleen of sokoto gudali cattle:

A Histomorphological study

*1Bello, A., 1Onu, J. E., 1Fawaz, A. M., 1Hena, S. A., 1Sonfada, M. L., 2Umaru, M. A., and 1Shehu, S. A.

1Department of Veterinary Anatomy, Usmanu Danfodiyo University, Sokoto, Nigeria.

2Department of Theriogenology and Animal production, Usmanu Danfodiyo University, Sokoto, Nigeria.

*Corresponding author E-mail: abccrcfge28@gmail.com.

Received 3 October 2016; Accepted 28 November, 2016

An anatomical study was conducted on the development of

cattle spleen using standard histomorphological methods.

Fifteen Sokoto gudali cattle fetuses obtained from Sokoto

metropolitan abattoir at different gestational ages were

used for the study. The fetuses were weighed and grouped

according to their gestational ages using their crown-

vertebral-rump length. Gross observation of the organ

shown that, the spleen was observed to be elongated, with

flat surfaces and rounded at the apices in all the stages of

development. At first trimester, the spleens were seen as

smooth massive organ, with almost uniform width and

thickness throughout the length. They were uniformly

pinkish, no definite shape and no visual regional

boundaries. At second trimester, the spleens were observed

to have taken the normal shape of adult cattle with definite

shape and visual regional boundaries (base, body and

apex). Biometrically, the weight of the foetuses, the crown –

vertebral – rump – length, weight of the spleen, length of

the spleen, width of the spleen and volume of the spleen

were found to be increasing progressively with

advancement in gestational age (first trimester to third

trimester). Histological observations showed that the

spleens had variable shape and size of the red pulp, white

pulp, trabercular connective tissue and capsular thickness

depending on the stage of development. A special feature of

connective tissue inter-digitations of variable sizes into the

parenchyma was observed along the whole surface at 2nd

and 3rd trimester. Based on the findings in the study, cattle’s

spleen had little/few variations with true ruminant and

much similarity with so many domestic animals in terms of

development.

Key words: Histomorphometry, Cattle, Gudali, Spleen,

Prenatal development.

INTRODUCTION Cattle, common term for the domesticated herbivorous mammals that constitute the genus Bos, of the family Bovidae, and that are of great importance to humans because of the meat, milk, leather, glue, gelatin, and other items of commerce they yield. Modern cattle are divided into two species: B. taurus, which originated in Europe and includes most modern breeds of dairy and beef cattle, and B. indicus, which originated in India and

is characterized by a hump at the withers (Atoji et al., 1998). The latter are now widespread in Africa and Asia, with lesser numbers imported to North America (primarily in the southern United States), Central America, and northern and central South America (Bello et al., 2012).

The spleen develops in the dorsal mesentery of the stomach (dorsal mesogastrium). The spleen arises from cells of the mesentery, which migrate into the plane

Bello et al. 10

between the layers of the mesentery. The mesentery covering the spleen becomes the visceral peritoneum of the spleen (Agungpriyono et al., 1995; Atoji et al., 1998; Bello et al., 2012). The mesentery between the spleen and the gut tube becomes the gastrosplenic ligament. The mesentery between the spleen and the dorsal body wall becomes the splenorenal ligament (most of which subsequently fuses to become parietal peritoneum).The Spleen is the largest lymphoid organ in the body, and the most important organ of immunological defense for blood invasion (Erdogan and Alan, 2012a; Franco et al., 1993). Most available reports on the histomorphological features of the spleen in humans (Dyce et al., 2002; El-Wishy et al., 1981), buffalo (Ateş et al., 2013; Banks, 1992), sheep (Kurtul and Atalgm,(1998); Salehi et al., 2010; Shao et al., 2010), goat (Dyce et al., 2002; Erdunchaolu et al., 1999; Igbokwe and Okolie, 2009; Sonfada, 2008; Bacha and Linda, 2000), rats (Getty, 1975; Kobayashi et al., 2005; Kumar et al., 1998; Wilson, 1995), camel (Bello et al., 2012a; Kurtul and Atalgm, 1998) and Llama (Agungpriyono et al., 1995; Bello et al., 2012a; Bello et al., 2012b; Zidan et al., 2000a) were entirely concerned with structures of adult, thus there is paucity of information about the prenatal development of cattle spleen in this region of the world.

In this research, gross changes, morphometric changes and histological differentiation involved in the development of the spleen of one-humped cattle will be described. The study will also add to the existing information on the morphometric analysis in cattle’s. Therefore this investigation was aimed at examining the gross and histological orientation of the spleen of Sokoto gudali cattle. MATERIALS AND METHOD The study was carried out on 15 fetuses of the gudalian cattle collected from the metropolitan abattoir, Sokoto at different gestational ages. The collected foetuses were then taken to the Veterinary Anatomy laboratory of Usmanu Danfodiyo University; where the weight and age of the fetus were determined. The fetal body weight was measured using electrical (digital) weighing balance for the smaller foetuses and compression spring balance (AT-1422), size C-1, sensitivity of 20kg X 50g in Kilogram for the bigger foetuses. The approximate age of the foetuses was estimated by using the following formula adopted by El-wishy et al. (1981) and Bello et al. (2012a). GA = (CVRL + 23.99)/0.366; Where GA is age in days and CVRL is the Crown Vertebral Rump Length. Fetuses below 130 days were designated as first trimester, 131- 260 days as second trimester and 261 –

390 days as third trimester (Bal and Ghoshal,1972). Crown Vertebral Rump Length (CVRL) was measured (cm) as a curved line along the vertebral column from the point of the anterior fontanel or the frontal bone following the vertebral curvature to the base of the tail. Based on this, fetal samples were divided into 3 main groups as described by Bello et al. (2012b).

The foetuses were then placed on a dorsal recumbency and a caudo-lateral skin incision was made at the thoraco-abdomino-pelvic region, to expose the entire digestive tract. The organs were examined in situ and exteriorised. The length, weight, width and volume of each organ were measured using ruler and thread, weighing balance, and vernier callipers, respectively. The length was taken from the cranial pole (apex) to the caudal pole (base) along the longitudinal axis while the width was taken as the distance between the two lateral aspects of the spleen, measured at the apex or the tip, mid-length (body) and root with vernier calliper, metric ruler for bigger spleen and recorded in millimeter. The Volume was determined by water displacement technique (Archimedes principle). The organs were weighed using an electronic weighing (Mettler® balance P1210, Mettler Instruments AG, Switzerland) with a sensitivity of 0.01 g. The data obtained were expressed as mean ± standard error of the mean (mean ± SEM).

Histological samples were taken as 1 cm2 thick,

collected and fixed in 10% formalin solution. After fixation was achieved, the tissue sample was processed for paraffin blocks preparation. The sections of 5 µm were subjected to haematoxylin and eosin for routine morphology (Bello et al., 2012a). The standard sections were examined under light microscope and micrographs taken using Sony digital camera(x5) with 12.1 mega pixel. RESULTS AND DISCUSSION From the fifteen (15) samples used for the study, five (5) belongs to the first trimester, five (5) from the second trimester and five (5) belongs to the third trimester. A total of ten (10) foetuses were males and five (5) were females (Table 1). The spleen of cattle was developed as an elongated and dorsoventrally flattened organ along its cranial two-thirds on the dorsal aspect of the stomach between the craniodorsal compartment and the caudodorsal compartment of the stomach from first trimester to third trimester with a rounded apex and a well-developed hilus (Figure 1). This finding is in agreement with the previous studies of Qayyum et al., (1991) on Llama; (Erdogan et al., 2012b) and (Kocak et al., 2011) on Bacterian cattle , (Dyce et al., 2002) on sheep; abundant as in ruminants (Qayyum et al., 1988) and (Kurtul and Atalgm, 1998); (Bal and Ghoshal, 1972) on Horse; (Iwasaki, 2002) and (Shao et al., 2010) on Sheep and in mouse, rat and hamster by Kobayashi et al.

Direct Res. J. Vet. Med. Anim. Sci. 11

Table 1. The biometric parameters of the foetuses and the spleen of cattle at various gestational age. Trimester Sample size (n) Mean CRVL (cm±SEM) Mean fetal weight (kg±SEM) Mean spleen weight (g±SEM)

1st

5 10.90±4.26 2.00±1.16 0.50±0.08

2nd

5 28.7±5.66 5.50±2.18 15.20±3.05

3rd

5 51.00±3.03 9.00±3.05 75.10±7.03

Table 2. The morphometrical parameters of the spleen of cattle at various gestational ages.

Trimester Sample size (n) Male female Mean length (cm ± SEM)

Mean breath (cm ± SEM)

Mean volume (cm3 ± SEM)

1st

5 2 3 2.60±0.04 1.00±0.30 5.50±0.08

2nd

5 3 2 4.00±0.62 1.70±0.14 24.20±0.05

3rd

5 3 2 10.00±0.12 3.90±0.40 51.10±0.03

Figure 1. Photomicrograph of Cattle Spleen (First Trimester) showing undifferentiated zones of white pulp (A) and Red pulp (B) cells with premature connective tissue fibers (Red arrow) H&E x200.

(2005) and Erdunchaolu et al. (2001).

This research showed that with advancement in gestational age, there were corresponding increases in the morphologic parameters. As observed in this work, the spleen is a dark red to blue black organ located in the left crainial abdomen which is flat and oblong in shape

and it is narrow as it extend in sheep and goat but has crescent shape in the cattle. The spleen was observed to be present from the differentiation of the digestive tract in the dorsal mesogastrium and as the stomach rotates during development the spleen comes to occupy the left crainial abdomen. Though grossly, the spleen present

Bello et al. 12

Plate 1. Photograph showing cattle foetus at 1

st trimester with

transparent abdominal wall (blue arrow) and rudimentary ear canal opening (black arrow) X 75.

Plate 2. Photograph showing cattle fetus at 2

nd trimester with thick

prominent skin (green arrow) and hair on the upper eyelid (black arrow) and head region X 75.

different shapes in different species of animals, its colour is usually dark red in most species. The spleen is enclosed in a capsule of fibrous and elastic tissue that extends into the parenchyma as traberculae. This finding is in agreement with the previous studies of Qayyum et al., (1991) on Llama; (Erdogan and Alan, 2012a) and (Kocak et al., 2011) on Bacterian camel, ( Dyce et al., 2002) on sheep; abundant as in ruminants, (Qayyum et al., 1988; Kurtul and Atalgm, 1998; Bal and Ghoshal,1972) on Horse; (Iwasaki, 2002) and (Shao et al., 2010) on Sheep and in mouse, rat and hamster by Kurtul and Atalgm, (1998) and (Erdunchaolu et al., 2001) (Plates 1,2 and 3).

Biometrically, the weight of the fetuses were observed to be ranging from 0.18±0.05 at first trimester to 21.70±7.28kg at third trimester, the crown – vertebral – rump – length were found to be 15.75±4.42 at first trimester to 94.00±2.83cm at third trimester and weight of the spleen were found to be 0.50±0.08g at first trimester to 110.10±7.03g at third trimester (Table 1). The observed biometric study of the foetuses and the spleen

were found to be progressively increasing with advancement in gestational ages. The weight of the fetus and spleen were found to increase significantly with advancement in gestational ages. This is in accordance with the work of Sonfada, (2008) and Bello et al. (2012b) who observed that there was an increase in body weight across the trimesters in the fetus with advancement in pregnancy and that of Wilson, (1995) that there was obvious body weight changes on mice which seemed to increase with age.

From the observed length, breath and volume, the increment was in accordance with Bello et al. (2012a) who showed that there was increase in the length, diameter, breath and volume of the digestive tract of cattle with increase in gestational ages (Table 2). This is also in line with Franco et al. (1993); Bal and Ghoshal, (1972) and Zheng and Kobayashi, (2006) on their works on digestive tract of bovine, porcine and caprine species respectively. In cattle, the spleen has a crescent shape and the mean length in first, second and third trimesters were 3.70 ± 0.05 cm ± SEM, 5.00 ± 0.29 cm ± SEM and

Direct Res. J. Vet. Med. Anim. Sci. 13

Plate 3. Photograph showing cattle fetus at 3

rd trimester with short densely distributed hair (whitish)

all over the body with very small areas of alopecia (black arrow). X 75.

Figure 2. Photomicrograph of Cattle Spleen (Second Trimester) showing clearly differentiated zones of white pulp (A) and Red pulp (B) cells with premature connective tissue fibers (Red arrow) H&E x200.

10.00 ± 0.94 cm ± SEM respectively. Histologically, the cattle spleen showed clearly differentiated zones of white pulp and red pulp but with premature traberculae connective tissue fibers. This advance with gestational age and thick traberculae connective tissue was seen before bath. In the first trimester, the spleen showed undifferentiated zones of white pulp and red pulp cells but with predominantly mesenchyme cells and thick connective

tissue fiber capsule as shown in (Figure 1). This is also in line with the findings of Franco et al. (1993), Bal and Ghoshal, (1972), Zidan et al. (2000a) and Atoji et al. (1998) on their works on digestive tract of bovine, porcine, canine and caprine species respectively. In the second trimester, there is clear differentiation of white pulp and red pulp cell with developing traberculae connective tissue fibers and thick fiber capsule as seen in (Figure 2). This finding is in agreement with the previous

Bello et al. 14

Figure 3. Photomicrograph of Cattle Spleen (Third Trimester) showing clearly differentiated zones of white pulp (A) and Red pulp (B) cells with mature traberculae connective tissue fibers connective tissue H&E x200.

studies of Qayyum et al., (1991) on Llama; Erdunchaolu et al. (2001) and Kocak et al. (2011) on Bacterian cattle, Dyce et al. (2002) on sheep. In the third trimester clear differentiation of white pulp and red pulp cells is seen with developed traberculae connective tissue as seen in (Figure 3). This finding is in agreement with the previous studies of Qayyum et al. (1991) on Llama; Erdunchaolu et al. (2001) and Dyce et al. (2002) on sheep; abundant as in ruminants Qayyum et al. (1988), Kurtul and Atalgm, (1998), Bal and Ghoshal, (1972) on Horse; (Iwasaki, 2002) and (Shao et al., 2010) on Sheep and in mouse, rat and hamster by Kurtul and Atalgm, (1998) and Iwasaki et al. (2002). However, in all the above studied, it was observed that the development of the spleen is completed at the 3

rd

trimester but maturity continues up to postnatal period. Conclusion Base on the above finding, the result shown that the development of spleen was found to be in succession i.e from the stages of ordinary mesenchyme cells with undifferentiated zones of white and red pulp to the stage of clearly differentiated zones of white pulp and Red pulp cells with developed traberculae connective tissue, well developed capsular connective tissue, with continous maturity at post-natal stage. These were seen with advancement in gestational age from first trimester

through second trimester and to third trimester. Specialization of the lingual muscles also followed the same trend of development, from immature unorganised muscle fibres to inmature organised fibres and finally matured organised differentiated muscle, with continous maturity at post-natal stage.

Authors` declaration We declare that this study is an original research by our research team and we agree to publish it in the journal.

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