Lebanese Science Journal, Vol. 18, No. 1, 2017
122
HISTOLOGICAL ALTERATIONS IN KIDNEY
AND LIVER OF LABORATORY MICE
FOLLOWING INTRAMUSCULAR INJECTION OF
MONTEVIPERA BORNMUELLERI (WERNER,
1898) VENOM
Eliane kossaifi1, Souad Hraoui-Bloquet2, Ryiad Sadek3, Ziad Fajloun4 , Claudine
Accary4 and Walid Hleihel5
1Biology
Departement, USEK, Kaslik-Jounieh, Lebanon
of Biology, Faculty of Sciences, Section II, Lebanese University, (Fanar),
B.P. 90656 Jdeidet El Maten, Lebanon
3Departement of Biology, American University of Beirut Lebanon
4Department of Biology, Faculty of Sciences, Section III, Lebanese University, El Kobeh,
Lebanon
5Faculty of Medical Sciences, USEK, Kaslik-Jounieh, Lebanon
sdbloquet@yahoo.com
2Département
(Received 10 February 2017 – Accepted 12 April 2017)
Abbreviations: LD50: Lethal Dose to kill 50% of the test population; IV: Intra venous; IP:
Intra peritoneal; SC: Subcutaneous; IM: Intra muscular; C: Capillary; Bs: Bowman’s space;
Pt: Proximal tubule; Dt: Distal tubule; PLA2: Phospholipase A2; LAAO: L-amino acid oxidase
ABSTRACT
Kossaifi, E. Hraoui-Bloquet, S. Sadek, R. Fajloun, Z. Accary, C. and Hleihel. W. 2017.
Histological alterations in Kidney and Liver of laboratory mice following intramuscular
injection of Montevipera bornmuelleri (Werner, 1898) Venom. Lebanese Science Journal,
18(1): 122-135.
Histopathological changes after bites of the Montivipera bornmuelleri viper, endemic
to high altitudes areas of Lebanon, have not yet been investigated. Our study focuses on
histological changes and irreversible damages in the kidney and liver of white laboratory
Balb/c mice that received an intramuscular injection of lyophilized venom diluted in saline
solution (NaCl 0.9%). Different venom concentration doses ranging from 0.25 mg/kg to
15mg/kg in a total injection volume of 100µl was injected intramuscularly (IM) into 5 groups
of mice. After dissection, observations showed no macroscopically identifiable damages in any
of the organs studied. However, tissue samples from the liver and the kidney were obtained for
histological studies at various time intervals following the venom injection. The histological
study was carried out using the Bouin solution (fixing bath), followed by dehydration in alcohol.
Paraffin-embedded sections were stained using hemalun-easine. Tissues from 6 control mice,
http://dx.doi.org/10.22453/LSJ-018.1.122135
National Council for Scientific Research – Lebanon 2016©
lsj.cnrs.edu.lb/vol-18-no-1-2017/
Lebanese Science Journal, Vol. 18, No. 1, 2017
123
which received only an injection of saline solution, were also examined. The common
histological alterations observed microscopically in the liver and the kidney were: pycnotic
nuclei, necrosis, vacuolization, cytoplasmic destruction, edema, hemorrhage and congestion of
blood vessels. Moreover, inflammatory infiltration of lymphocytic cells was observed in the
prevascular regions of both organs. The histological changes observed appeared within 1h and
intensified with time. These changes were proportional to the dose of the venom injected into
the laboratory mice. Our results on what happens in the kidney and liver of mice after injecting
the venom of M. bornmuelleri had many similarities with the effect of venom of other vipers (e.
g. Daboia palestinea, Bitis ssp, Bothrops moujenis etc…) observed by others authors.
Keywords: Montevipera bornmuelleri, lyophilized venom, intramuscular injections,
liver, kidney.
INTRODUCTION
M. bornmuelleri is endemic to Lebanon (Werner, 1938) and only found in Lebanese
mountains at altitudes above 1800m (Hraoui-Bloquet et al. 2002). It is now listed as endangered
by IUCN (2006) based on its limited and patchy geographic range estimated to be less than 20
000 km2. Moreover, its population is highly fragmented in the mountains of Lebanon. Studies
concerning M. bornmuelleri are very rare. An ecological study with preliminary biochemical
characterization of the viper’s venom was conducted by Hraoui-Bloquet et al. in 2012. Other
recent pioneer studies concerning proteomic analysis and biological properties of the M.
bornmuelleri venom were conducted (Rima et al., 2013; Accary et al., 2014a; Accary et al.,
2014b; Accary et al., 2015).
Lethality determinations were reported for Montivipera (Vipera) bornmuelleri and
compared to Montivipera (Vipera) latifii by Weinstein and Minton (1984).
Immunoelectophoretic profiles indicated a close antigenic relation between venoms of these
species with Daboia palestinae and Bitis spp.
Damages caused in tissues from the snake envenomation depend on the type and
amount of venom injected, as well as on the species because not all venoms have the same
composition. Those damages rest also in the susceptibility of the tissue studied for a particular
component of the venom (kamiguti et al., 2000). Interestingly, viper venom affects the kidney
tissues more frequently than other snakes. Renal alterations are also very common following
viper bites (Sheriff, 1983). These alterations are more prominent when the venom contains
hemotoxins
and
vasculotoxins.
A
histological
study
demonstrated
that
alterations/damages/outcomes in renal tissues are very common after viper bites (Soe et al.,
1993). The changes have been observed in all renal structures. Biochemical and clinicopathological changes were induced by Bungarus caeruleus venom in the rat model (Kiran et
al., 2004). It was shown that changes observed in the kidney after the administration of snake
venom include degeneration, necrosis and regeneration of renal tubular epithelial cells,
interstitial edema, cellular infiltration, arteritis, thrombophlebitis, congestion, infarction and
cortical necrosis (Kiran et al. 2004, Stiprija et al. 2006). Histological and functional renal
alterations, caused by Bothrops moujeni snake venom in rats,asserted the lymphatic
contribution to the systemic absorption of venom toxins from the tissues (Boer-Lima t al.
1999). Therefore, studies of potential effects of snake venoms on the structure and function of
lymphatic vessels, might have implications for the pathogenesis of edema and the absorption
of venom by the tissues (Gutierrez, et al. 2003).
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124
Changes in the liver following snake bites were reported in studies conducted by De
Silva et al. (1992) and Kumaranayake (1971). Both studies observed congestion, petechial
haemorrhages, microvescicular fatty change, hydropic degeneration,and necrosis of
hepatocytes. The above-mentioned observations are in close accordance with the findings
reported for Vipera berus and Daboia (Vipera) palaestinae (Tu, 1977).
Jarrar (2011) studied the histological alterations and biochemical changes in the liver
of sheep following Echis colora envenomation. This venom elevated glucose, , aspartate amino
transferase (AST), alanine aminotransferase (ALT), triglyceride and total bilirubin, while
cholesterol was reduced. The histological alterations detected, were pyknosis, karyorrhexis,
cytoplasmic vacuolation, necrosis, fatty changes, hepatocytes atrophy, sinusoidal dilatation,
kupffer cell activation, amyloidosis, portal vein thrombosis, partial glycogen depletion and
hepatic architecture distortion. These findings revealed that E. Colora venom produced
biochemical changes and histological alterations in the liver, which may have a severe effect
on the functions of this organ.
Nanayakkaraetal. (2009) described the histological changes in liver, kidney and brain
tissues following intramuscular administration of the venom of Bungarus ceylonicus and
Bungarus caeruleus. Histopathological changes relative to congestion, inflammation and
necrosis were also observed microscopically in these tissues. These changes were proportionate
to the dose of venom.
In this study, we aim to identify the histological changes in liver and kidney tissues
following an intramuscular injection of M. bornmuelleri venom, at different concentrations,
into white mice (Balb/c).
MATERIALS AND METHODS
Venom
Venom was extracted from 6 adult specimens of M. bornmuelleri caught in Jabal
Sannine and Jabal Makmel (two mountains located in Lebanon). Stock solution* of the venom
were prepared immediately before the experiments. This was done using 25 mg of fresh venom
in 5 ml of a phosphate buffered saline solution at pH 7.
The LD50 of M. bornmuelleri previously determined by Weinstein S. and Minton S.
(1984) corresponds to 0.6 mg/kg in intra venous (I.V.) injection and 1.9 mg/ kg in intra
peritoneal (I.P.) injection and 6.2 mg/kg in subcutaneous (S.C.) injection. Here, we adopted the
LD50 obtained by intramuscular (I.M.) injection of the fresh, rather than lyophilized venom
and determined to be 5.39 mg/ kg for mice weighing between 25 and 30 g (Abi-Rizk et al.,
2017).
Animals and treatments
Six groups of white laboratory mice (Balb/c), 2 to 3 months old, weighing 25-30 gr were used.
Aliquots of 0.2 ml of the venom at different concentrations (Table 1) were injected
intramuscularly ( I.M.) into the left thighs of mice. The intensity of venom used was based on
125
Lebanese Science Journal, Vol. 18, No. 1, 2017
the LD50 (5.39 mg/kg) values for M. borrnmuelleri. A control group (or group No 1) was
injected with 0.2 ml of phosphate buffered saline solution at pH 7.
TABLE 1
Different Concentrations and Volumes Injected in Mice by IM of the Venom Dilutions
of M. bornmuelleri Venom
Group
Numbers
Solution injected
1
Injected volume
Individual
Number
6 (control)
Phosphate
0.2ml
buffered saline
2
Stock solution*
0.2 ml
16
3
8 mg/kg
0.2 ml
16
4
4 mg/kg
0.2 ml
16
5
1.6 mg/kg
0.2 ml
16
6
0.8 mg/kg
0.2 ml
16
Stock solution* see paragraph venom in Materials and Methods
Action time
24h
1h-3h-5h-24h
1h-3h-5h-24h
1h-3h-5h-24h
1h-3h-5h-24h
1h-3h-5h-24h
Experimental protocol
After 1h, 3h, 5h and 24h following the administration of venom to the mice, livers
and kidneys were removed and fixed in Bouin solution for 24 h. The tissues were then washed
witch tap water, dehydrated in a graded ethanol series (70%, 90% and 100%) and embedded in
paraffin at 60 ºC.
Histopathologic analysis
5 µm thick paraffin sections were prepared and stained with hematoxylin and eosin
and then assessed under light microscopy. Histological changes were scored according to a
predetermined system given in table 2.
Results
Some mice died directly after injection of the venom while others died sometime
before dissection (table 2). These mices were eliminated from the study.
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TABLE 2
Venom Concentrations, Number of Mice Injected and Number of Dead Mice Observed
during the Experiments
Serial numbers
S1 & S6
ST3 & ST6
Solution
concentrations
Stock solution*
1/5 :
Number of mice
injected (IM)
16
ST1
S2 & S4
dilution
mg/kg
8
16
S7 & S11
dilution 1/25 : 1.60
mg /kg
16
Number of dead mice
and surviving time
2 mice ( just after solution
injection), 2 mice (30 min
after solution injection)
1 mice ( just after solution
injection) 2 mice (15 min
after solution injection)
2 mice (just after solution
injection)
All envenomated mice presented the following visual observations/symptoms in the
region of the injection: hemorrhage, edema, dermonecrosis and myonecrosis reaction after
intramuscular (I.M.) of the venom. Microscopic observations of histological sections of livers
and kidneys from all envenomated mice presented many common histopathological changes
like necrosis, pyknotic nuclei, edema, congestion of blood vessels, tearing of the vessel walls,
bleeding and inflammations (inflammatory infiltrates consisted of lymphocytes).
Histopathological changes, observed due to the administration of this venom, appeared within
1h in liver and in kidney.
Histopathology of liver: Changes observed in histological tissues sections
Histopathological changes namely vessel congestion, sinusoidal dilatation,
inflammation, necrosis, suffering damage (degradation or degeneration) and depletion of
hepatocytes, hepatic architecture distortion and edema were microscopically observed in the
liver tissues. These findings revealed that M. bornmuelleri venom altered the histology of the
liver of the envenomated mice. Accordingly, these alterations might severely affect the
functions of this organ. They were acutely observed in livers of mice that received the stock
solution or the dose concentration of 8 mg/mice kg. Moreover, congestion of the portal vein
was seen in almost all liver tissues from 1h to 24 h post-injection (Fig.1B ).
Cg
Ed
A: healthy liver section showing the
hepatocytes in good shape (
)
B: liver section after 1h of venom
injection showing congestion (
Cg)
and an edema (
Ed) of the portal
vein.
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Lebanese Science Journal, Vol. 18, No. 1, 2017
Inf
C: liver section after 1h of venom injection
showing the inflammatory infiltration
(
Inf)
Cv
At
Nc
D: liver section after 1h of venom
injection showing the necrosis
hepatocytes (
Nc)
E: liver section after 1h of venom
injection showing the diseased
hepatocytes
with
cytoplasm
vacuolation (
Cv) and hepatocyte
atrophy
(
At)
Figure 1. Five pictures taken after staining and microscopic observations of five liver
sections. A: section of the liver of healthy mice not injected by the venom; B, C, D and E:
sections of the livers of mice after 1 hour of IM injection of Montivipera bronmuelleri
venom showing congestion, edema, inflammatory infiltration, necrosis hepatocytes and
diseased hepatocytes respectively.
Inflammatory infiltration (Figure 1D) was seen in the perivascular region of the liver.
Inflammatory infiltrates were confined to the periportal and perivenular regions of the liver,
and appeared in tissues observed from 1h to 24h post-injection following the administration of
the venom (Table 3). Liver necrosis (Figure 1E) was a consistent finding following the venom
injection. Karyopycnosis of some hepatocytes was evident namely in the necrotic hepatocytes.
A variable degree of hepatocyte cytoplasmic vacuolation was detected (Figure 1E), where its
severity was associated with necrosis. Hepatocytes suffering and atrophy were also observed.
It is noteworthy that edema was observed in all cases of envenomated mice.
These sections were compared to those from control mice that were not injected with
the M. bornmuelleri venom.
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Lebanese Science Journal, Vol. 18, No. 1, 2017
TABLE 3
Histopathological Changes Observed in Liver Tissues after IM Injection of Different
Doses of M. bornmuelleri Venom (Stock Solution; 1/5=8mg/kg; 1/10=4mg/kg;
1/25=1.6mg/kg; 1/50=0.8mg/kg).
Number of
samples
3
6
12
13
16
Doses
(mg/Kg)
Control
Saline
solution
Stock solution
8 mg/kg
4 mg/kg
14
1.6mg/kg
16
Time
congestion
Inflammation
edema
Hemorrhag
e
-
Suffering cells
-
necrosi
s
-
24h
24h
2+
1+
1h
1h - 5h
1h - 5h
12 ++
13 ++
14 ++
10 +++
9 ++
12 +
12 +
11 +
16 ++
12 +++
12 +++
16 +
10 ++
9 ++
10 +
12 +++
13 ++
16 ++
8+
14 +
13+
13+
1h 12 +
12 +
12 +
14 +
24h
0.8mg/kg
1h 13 +
13 +
13 +
13+
24h
The sign (+) Indicates the Presence of the Anomaly/Histopathology.
+ = Moderate; ++ = High; +++ = Very high; - = Absent
Histopathology of kidney: Changes observed in the tissues sections of the Kidney
Histopathological changes namely congestion, inflammation and necrosis were
observed microscopically in the tissues of the kidney. Peritubular capillary and glomerular
congestion were also seen in most tissue samples of the kidney (Figure 2B). On the other hand,
inflammatory infiltration, mainly lymphatic (Figure 2C), was found in the perivascular regions
of the kidney. Marked changes and severe morphologic disturbances were observed in the
glomeruli after venom injection. Histological examinations of the kidney showed wide
Bowman’s spaces, filled with debris of cells, and damaged glomeruli (Figure 2D and 2E).
Severe morphologic disturbances or degenerations, especially in the distal tubules,
were unveiled. Degenerative changes were also seen in the proximal tubules after
envenomation. These changes consisted of the loss of the proximal brush border, the
cytoplasmic vacuolation, or in some tubules the degeneration and desquamation of the necrotic
cells (Figure 2F). Cell debris and casts resulting from necrosis were observed in the collecting
ducts and in the lumen of the proximal and distal tubules (Figure 2F). The nuclei of the various
proximal tubular cells often showed pyknosis with clumped chromatin material. In some
tubules, the renal epithelium was completely necrotic, whereas the basal membrane was either
intact or disrupted by tubular necrosis (Fig.2F). The distal tubules, nephron loops, and
collecting ducts had swollen lumens. In some areas, there were disruptions of both tubular walls
(Figure 2F) and peritubular capillaries walls, indicating hemorrhage (Figure 2D). Additionally,
hyaline casts, red blood cells and sloughed cellular debris filled the lumens. Necrotic and
damaged areas were present in tubules of the cortico-medullary regions. All these damaged
tissues were compared to healthy kidney section of mice not injected by the M. bornmuelleri
venom (Figure 2A).
-
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Lebanese Science Journal, Vol. 18, No. 1, 2017
Ed
Cg
A: healthy kidney section showing the good
quality of epithelial cells of different tubules
(
)
B: kidney section after 1h of venom
injection showing congestion (
Cg)
and an edema (
Ed) at the cortical area
Bd
Inf
Hm
C: kidney section after 1h of venom injection
showing the inflammatory infiltration
(
Inf)
D: kidney section after 1h of venom
injection showing the Bowman’s spaces
dilated (
Bd) and disruption of
capillary wall with hemorrhage (
Hm)
Cd
En
E: kidney section after 1h of venom injection
showing the cell debris in the dilated lumen
of the Bowman’s space (
Cd) and damage
of its basal membrane
F: kidney section after 1h of venom
injection showingrenal epithelium damage
of the epithelium and basal membrane of the
distal tubule (
En).
Figure 2. Six pictures taken after staining and microscopic observations of six kidney
sections. A: section of the kidney of healthy mice not injected by the venom; B, C, D, E
and F: sections of the kidney of mice after 1 hour of IM injection of M. bronmuelleri
venom showing congestion, edema, inflammatory infiltration, Bowman’s space
dilatation, necrosis epithelial cells of tubules respectively.
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Lebanese Science Journal, Vol. 18, No. 1, 2017
TABLE 4
Histopathological Changes Observed in Kidney Tissues after IM Injection of Different
Doses of the M. Bornmuelleri Venom (Stock Solution; 1/5=8mg/kg; 1/10=4mg/kg;
1/25=1.6mg/kg; 1/50=0.8mg/kg).
Number
samples
Doses (mg/Kg)
Time
Mice
congestions
3
Control
24h
-
6
12
13
16
14
Saline solution
Stock solution
8mg/kg
4mg/kg
1.6mg/kg
16
of
with
Inflammation
edema
hemorrhage
Glomeruli
changes (C &
Bs)
-
-
-
-
Necrosis
&Suffering
cells in (Pt &
Dt)
-
12 +++
13 +++
16 +
8+
12+++
13 +++
16+
10+
-
-
24h
1h
12++
12++
12 ++
10+
1h - 5h
13++
13++
10 ++
8+
1h - 5h
14++
13 ++
16 ++
10+
1h 10+
6+
8+
24h
0.8mg/kg
1h 12+
24h
The Sign (+) Indicates the Presence of Anomaly.
C: capillary, Bs: bowman’s space; Pt: proximal tubule; Dt: distal tubule
+ = Moderate; ++ = High; +++ = Very high; - = Absent
CONCLUSION
Severity of the envenomation depends on several factors such as: the snake species,
the bitten animal, the host animal size, the amount of venom injected (Latifi, 1995) because of
the large variability in the venom components.
Histological changes followingM.
bornmuelleribites are not yet documented. We chose to evaluate the liver and kidney condition
after IM injection of M. bornmuelleri venom since, they are the two first organs which are
damaged in the body. The histopathological changes observed were of an acute toxic insult and
the snake venom significantly affected the function of these organs (Nanayakkaraetal., 2009).
We have demonstrated that the administration of the M. bornmuelleri venom resulted in
congestion, inflammation, edema, hemorrhage and necrosis in the liver and the kidney at all
concentrations only 1 hour after the injection.
Lower molecular weight toxins in krait venom are rapidly absorbed and quickly
distributed through the blood stream (De Silva and Ranasinghe, 1983). The first tissue changes
were detected in the brain, kidney and liver 1 hour after injection. Accordingly, toxic changes
would appear early in the target organs. Our results demonstrated that the M. bornmuelleri
venom causes irreversible histological damages in the kidney and liver of mice 1 hour after
injection. Histopathological changes caused by toxins present in the venom of M. bornmuelleri
that resemble those observed by Nanayakkara et al., (2009) who reported many changes like
congestion, inflammation and necrosis in kidney and liver caused by the krait venom of
Bungarus.
Viper venom, which is rich in phospholipases, causes intensive tissue damage in
many organs. Phospholipases present in viper venom are toxic (Fry et al., 2003). These enzymes
Lebanese Science Journal, Vol. 18, No. 1, 2017
131
hydrolyze phospholipids in the cell membrane and disturb the cell membrane activity (Iwanaga
and Suzuki, 1979). Here, we can consider that the histopathological manifestations, mainly
inflammatory and necrotic in nature, could have occurred in response to the membrane damage
caused by the action of these phospholoipases present in M. bornmuelleri venom. PLA2
enzymes induce edema and has been previously described in several snake venoms (Ali et al.,
1999). Thus, edema was observed in all liver and kidney tissues of mice injected with different
doses of M. bornmuelleri venom during our experiments. This can be explained perhaps by the
presence of LAAO in the venom (Rima et al., 2013). The LAAO is widespread in nature, where
snake venoms are apparently the richest sources. Several authors have reported that the LAAO
was able to induce edema in the lung and is in some cases accompanied by slight bleeding
(Stabelietal., 2004; Izidoroetal, 2006). Other previous studies demonstrated that hemorrhage,
hypotension, inflammation could be caused by serine-proteases, PLA2, metallo-proteases and
lecitines of type C (Kardoug, 2002), all of which were detected in the M. bornmuelleri venom
(Accaryetal., 2014a).
The purified PLA2 reveals several biological effects including pro-inflammatory,
antimicrobial, anticoagulant and hemolytic activities. Serine-proteases called SVMPS have
been isolated primarily from the venoms of viperidae snakes. Therefore, these thrombin-like
enzymes are widely distributed in venoms of several kinds of snakes. They affect many
physiological processes. SVMPs being hemorrhagic, induce edema and increase vascular
permeability (Rodrigues etal., 2001; Fernandesetal., 2006), thus contributing to the
inflammatory response which is characteristic of envenomation by viperidae (Teixeira et al.,
2003). This effect depends not only on the hydrolysis of the components of the basal membrane
but also on the release of inflammatory mediators from the cells (Wei et al., 2003). SVMPs
produce a substantial inflammatory leukocyte infiltration associated with an increase in the
number of circulating leukocytes (Costa et al. ., 2002; Fernandes et al., 2006). Metalloproteases Zinc-dependents are abundant in viperidea venom (Francischi etal., 2000; Markland
and Swenson, 2013). Metallo-proteases SVMPs induce, in the injection region, hemorrhage,
edema,necrosis of the muscle, dermonecrosis, and a substantial inflammatory reaction. All
envenomated mice after IM injection of the M. bornmuelleri venom showed the same
alterations (Abi-Rizk et al., 2017).
The highest hemorrhagic activity is associated with SVMPs of the P-III class, that are
the more active,such asVaH4 isolated from the venom of Viperaammodytes (Sajevicet al.,
2013) and the VLH2 isolated from the venom of Macrovipera (Vipera) lebetina(Hanzetal.,
2010). SMVPs induced alterations in the endothelial cells and their basal membrane (Ohsakaet
al., 1973; Franceschiet al., 2000; Rodrigues etal., 2000; Rucavadoet al.1998; Gutierrez et al.
1995).
Nanayakkaraetal., (2009), who studied the effect of elapid venom on mice kidney,
described that the main route of excretion of toxins is via the kidney. This would explain the
presence of congestion and inflammation in this organ. These two phenomena were observed
in the kidney of mice after injection of M. bornmuelleri venom. However, absence of nephritic
agents in elapid venom, vasculotoxins and haemotoxins would be the reason for the lack of
necrosis in the kidneys. These two toxins present in viper venoms explain necrosis and
hemorrhage observed in the kidney and liver of the mice treated with M. bornmuelleri venom.
Boer-Lima, (1999) described the histopathological alterations in rat kidney caused by
Bothropsmoojeni snake venom. The injection of this venom led to acute tubular and glomerular
Lebanese Science Journal, Vol. 18, No. 1, 2017
132
changes compatible with acute renal failure such as a decrease in the glomerular filtration rate
and a sustained increase in tubular sodium rejection. Histological observations indicated
morphological abnormalities such as tubular swelling and glomerular damage. These damages
were largely resolved after 48h. More so, no resolved damages were observed in mice injected
by M. bornmuellei venom. The presence of mitotic tubule cells showed that epithelial
regeneration occured. Indeed, mitosis has been reported to coexist with areas of necrotic renal
parenchyma (Lieberthal and Levinsky, 1992). We also observed the phenomena of mitosis in
the kidney of mice envomated by M. bornmuelleri venom.
This study shows that the action of the venom of M. bormuelleri in small doses can
cause very serious damage to vital organs such as the liver and kidney. The determination of
the lethal dose shows that the toxicity of M. bornmuelleri venom is very high in comparison
with other species of vipers (Abi-rizk et al., 2017). For high doses, greater than or equal to lethal
degree (LD50) which is 5.39 mg / kg, the cellular damage observed is enormous in these two
organs. All cells appear to be destroyed and nonfunctional. This irreversible damage is observed
at the membrane, cytoplasmic and nuclear levels, causing the death of the animals injected.
This venom also acts on the vascular system of these organs causing capillary endothelial
ruptures and haemorrhages. The venom of M. bornmueller contains phospholipases,
metalloproteases, transferases, etc… (Hraoui-Bloquet et al., 2012, Accary et al., 2014a)
This study would be of importance in the characterization of possible proteins in M.
bornmuelleri venom, as well as in understanding their action in the process of development of
specific anti-venom. In addition, a further comprehension of the toxicology of M. bornmuelleri
is of local importance because of the spread of this species in Lebanon.
Acknowledgement
The financial support of the National Council for Scientific Research (CNRS-Lebanon) to this
study is highly appreciated.
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